Analysis of mutant parasites that have a defect in adhésion - Conférence du 6e édition du Cours international « Atelier Paludisme » - MATTEI Denise - France - dmm@pasteur.fr
1. The study aimed to investigate why clade A cyanopodoviruses are more virulent than clade B by examining whether they encode a thioredoxin (trx) gene, which increases replication rate.
2. PCR and sequencing were performed on genomic regions of interest from clade A and B cyanopodoviruses. Results showed that clade A viruses TIP28 and TIP33 contained homologs of trx and other genes, while clade B virus TIP41 contained a hypothetical protein gene.
3. The findings support the hypothesis that encoding of trx by clade A but not clade B viruses contributes to their higher virulence, through increasing replication
Cyclic di GMP signaling network in Legionella pneumophilaasslev
This presentation sums up a 3 years long research aimed to study the involvement of the ubiquitous bacterial cyclic di-GMP signaling network in Legionella\’s host infection and intracellular multiplication.
Coronin-1C and caveolin provide parallel pathways for trafficking Rac1 in migrating fibroblasts. Coronin-1C constitutively recycles Rac1 from the rear and sides of cells to maintain Rac1 levels. Caveolin endocytosis also removes Rac1 from the rear and sides, but targets it for degradation upon fibronectin engagement of syndecan-4. These redundant trafficking pathways facilitate sustained, polarized migration by defining membrane regions resistant to Rac1 activation once a chemotactic gradient dissipates.
This document summarizes a study on the localization and potential functions of two proteins, VDAC4 and TGrPE1, in the ciliated protozoan Tetrahymena thermophilia. VDAC4 encodes a porin protein that localizes to mitochondria and basal bodies, suggesting a role in mitochondrial-ciliary transport. TGrPE1 encodes a protein containing a GrPE1 domain that may regulate chaperone proteins through ATP/ADP exchange, and it localizes near mitochondria. The goal is to better understand the roles of these proteins in mitochondrial processes and protein folding in Tetrahymena.
The document discusses using surface entropy reduction (SER) mutations to improve crystallization of an RNA-Fab complex. Key points:
- SER mutations were identified on the Fab surface using software to reduce flexibility and promote crystal contacts. Mutations were incorporated using Kunkel mutagenesis.
- Mutant Fabs (Fab2SMA, Fab2SMS) were expressed at 3-4 mg/L and bound RNA similarly to the wild type Fab. Initial crystal hits were found for the mutant complexes.
- The SER approach aims to generalize to other RNA-Fab complexes, as demonstrated by binding of mutant Fabs to the VCIII riboswitch. Optimization of crystal hits is ongoing to improve diffraction resolution
JACS-CD38 localization using a fluorescent probeJonathan Shrimp
This document describes the development of a fluorescent small molecule probe called SR101−F-araNMN that can label the enzyme CD38 in live cells in a mechanism-based manner. The probe was used to investigate the cellular localization of CD38 in leukemia (HL-60 and K562) and lymphoma (Raji) cell lines. The results showed that CD38 is predominantly localized to the plasma membrane in Raji and RA-treated HL-60 cells, with very little intracellular CD38. Additionally, no CD38 expression was detected in K562 cells. This suggests that the major function of CD38 is to hydrolyze extracellular rather than intracellular NAD.
1) The study found that deletion of the protein cIAP1 in mice alters actin cytoskeleton organization, preventing the formation of filopodia in response to TNF.
2) cIAP1 was shown to directly interact with and regulate the activity of the small GTPase cdc42, which controls filopodia formation. Specifically, cIAP1 promotes the stabilization of cdc42 by facilitating its interaction with RhoGDIa in the cytosol.
3) TNF stimulation disrupts the interaction between cIAP1 and cdc42, allowing cdc42-mediated rearrangement of the actin cytoskeleton and formation of filopodia independently of NF-kB activation
This document summarizes a study investigating the roles of Rho and ADP-ribosylation Factor (ARF) GTPases in regulating phospholipase D (PLD) activity in human lung adenocarcinoma cells. The study used recombinant Sindbis viruses to express Clostridium botulinum C3 exoenzyme (which inactivates Rho) and a dominant-negative Rho mutant in the cells. Expression of C3 or the mutant Rho increased basal PLD activity and decreased phorbol ester-stimulated PLD activity. Bradykinin- or sphingosine-stimulated PLD activity, which had additive effects, was abolished by C3 or mutant Rho expression. Brefeld
1. The study aimed to investigate why clade A cyanopodoviruses are more virulent than clade B by examining whether they encode a thioredoxin (trx) gene, which increases replication rate.
2. PCR and sequencing were performed on genomic regions of interest from clade A and B cyanopodoviruses. Results showed that clade A viruses TIP28 and TIP33 contained homologs of trx and other genes, while clade B virus TIP41 contained a hypothetical protein gene.
3. The findings support the hypothesis that encoding of trx by clade A but not clade B viruses contributes to their higher virulence, through increasing replication
Cyclic di GMP signaling network in Legionella pneumophilaasslev
This presentation sums up a 3 years long research aimed to study the involvement of the ubiquitous bacterial cyclic di-GMP signaling network in Legionella\’s host infection and intracellular multiplication.
Coronin-1C and caveolin provide parallel pathways for trafficking Rac1 in migrating fibroblasts. Coronin-1C constitutively recycles Rac1 from the rear and sides of cells to maintain Rac1 levels. Caveolin endocytosis also removes Rac1 from the rear and sides, but targets it for degradation upon fibronectin engagement of syndecan-4. These redundant trafficking pathways facilitate sustained, polarized migration by defining membrane regions resistant to Rac1 activation once a chemotactic gradient dissipates.
This document summarizes a study on the localization and potential functions of two proteins, VDAC4 and TGrPE1, in the ciliated protozoan Tetrahymena thermophilia. VDAC4 encodes a porin protein that localizes to mitochondria and basal bodies, suggesting a role in mitochondrial-ciliary transport. TGrPE1 encodes a protein containing a GrPE1 domain that may regulate chaperone proteins through ATP/ADP exchange, and it localizes near mitochondria. The goal is to better understand the roles of these proteins in mitochondrial processes and protein folding in Tetrahymena.
The document discusses using surface entropy reduction (SER) mutations to improve crystallization of an RNA-Fab complex. Key points:
- SER mutations were identified on the Fab surface using software to reduce flexibility and promote crystal contacts. Mutations were incorporated using Kunkel mutagenesis.
- Mutant Fabs (Fab2SMA, Fab2SMS) were expressed at 3-4 mg/L and bound RNA similarly to the wild type Fab. Initial crystal hits were found for the mutant complexes.
- The SER approach aims to generalize to other RNA-Fab complexes, as demonstrated by binding of mutant Fabs to the VCIII riboswitch. Optimization of crystal hits is ongoing to improve diffraction resolution
JACS-CD38 localization using a fluorescent probeJonathan Shrimp
This document describes the development of a fluorescent small molecule probe called SR101−F-araNMN that can label the enzyme CD38 in live cells in a mechanism-based manner. The probe was used to investigate the cellular localization of CD38 in leukemia (HL-60 and K562) and lymphoma (Raji) cell lines. The results showed that CD38 is predominantly localized to the plasma membrane in Raji and RA-treated HL-60 cells, with very little intracellular CD38. Additionally, no CD38 expression was detected in K562 cells. This suggests that the major function of CD38 is to hydrolyze extracellular rather than intracellular NAD.
1) The study found that deletion of the protein cIAP1 in mice alters actin cytoskeleton organization, preventing the formation of filopodia in response to TNF.
2) cIAP1 was shown to directly interact with and regulate the activity of the small GTPase cdc42, which controls filopodia formation. Specifically, cIAP1 promotes the stabilization of cdc42 by facilitating its interaction with RhoGDIa in the cytosol.
3) TNF stimulation disrupts the interaction between cIAP1 and cdc42, allowing cdc42-mediated rearrangement of the actin cytoskeleton and formation of filopodia independently of NF-kB activation
This document summarizes a study investigating the roles of Rho and ADP-ribosylation Factor (ARF) GTPases in regulating phospholipase D (PLD) activity in human lung adenocarcinoma cells. The study used recombinant Sindbis viruses to express Clostridium botulinum C3 exoenzyme (which inactivates Rho) and a dominant-negative Rho mutant in the cells. Expression of C3 or the mutant Rho increased basal PLD activity and decreased phorbol ester-stimulated PLD activity. Bradykinin- or sphingosine-stimulated PLD activity, which had additive effects, was abolished by C3 or mutant Rho expression. Brefeld
This document characterizes a diversity-generating retroelement (DGR) in Legionella pneumophila that confers adaptive advantages by diversifying a target protein. The DGR contains all the core components found in other DGR systems and is capable of transferring DNA sequence from a template repeat to a variable repeat of the target gene ldtA, accompanied by adenine-specific mutagenesis. The target protein LdtA is anchored in the bacterial outer membrane with its variable C-terminal domain exposed on the surface, allowing it to generate massive protein diversity on the bacterial surface. Analysis of related DGRs in other L. pneumophila strains showed they target different carrier proteins but maintain similar variable domains, demonstrating the modularity and adapt
A high copy number screen of the trs23Δ99C mutantGabriel Bendavit
This document summarizes a study investigating the role of the TRAPP I subunit TRS23, a guanine nucleotide exchange factor for the Ypt1/Rab GTPase, in the secretion pathway of yeast. A high copy number screen was performed on a yeast mutant with a C-terminal deletion of TRS23 (trs23Δ99C) to identify suppressors of its thermosensitive phenotype. From screening 36,000 transformed yeast colonies, 37 colonies showed suppression. Sequencing identified the gene CIN5, involved in chromosome instability, as a potential suppressor. Further experiments, such as isolating and transforming CIN5 into the trs23Δ99C mutant, are needed to confirm its role in suppress
This document discusses using lambda bacteriophage as a platform for phage display. It describes the major capsid proteins gpE and gpD, which forms trimers and decorates the capsid surface. Mutants called IPDF (inhibited by gpD-fusion) and supIPDF (suppressor of IPDF) were isolated that affect plating efficiency in the presence of gpD-fusions. Sequencing showed the mutations were not in gpD or gpE, but likely other capsid proteins like gpC that interact with them. Future work aims to generate mosaic lambda display particles by co-expressing gpD and gpD-fusions from modified lambda lacking gpD to study capsid incorporation.
This document reports the 1H, 13C, and 15N backbone resonance assignments of the 214 amino acid human DGCR8core protein (residues 493-706) determined using heteronuclear NMR spectroscopy. DGCR8core contains two tandem double-stranded RNA binding domains (dsRBDs) separated by a flexible linker that are required for recognizing and binding pri-miRNA substrates during miRNA biogenesis. The NMR assignments provide a foundation for further investigating the dynamics and RNA-binding properties of DGCR8core in solution. Secondary structure analysis using chemical shift indices matches the seven alpha helices and seven beta strands observed in the crystal structure of DGCR8core.
AJS: a lethal weapon to combat with cancers Treatment of Human Cancers Using...gan-navi
This document describes a lentiviral vector called AJS2001 that can selectively silence the Cdc6 gene in cancer cells. Cdc6 is essential for initiating DNA replication, and silencing it blocks cancer cell proliferation. The vector was shown to inhibit growth of several cancer cell lines through Cdc6 knockdown, inducing senescence. Case studies are presented of cancer patients who recovered after treatment with injections of AJS2001 lentivirus, including patients with gastric carcinoma, brain and liver metastases from ovarian cancer, lung and larynx metastases from parotid carcinoma, and breast cancer.
This document summarizes a thesis project investigating the regulation of the anaerobic responsive transcription factor TdcA in Salmonella Typhimurium. The project aims to construct Salmonella strains with deletions or tags of genes involved in regulating tdcA expression, and to analyze the effects on tdcA transcription and protein binding to its promoter region. Specifically, the project will create ∆tdcA and tdcA-FLAG strains to identify members of the TdcA regulon, and use transcriptional analysis and chromatin immunoprecipitation to study how proteins like CRP, FNR, and H-NS regulate tdcA expression in Salmonella. Understanding TdcA regulation is important because it plays a role in Salmon
The document summarizes key concepts about the cell cycle, including:
1) The cell cycle consists of interphase (G1, S, G2 phases) and mitosis (M phase), with checkpoints between phases to ensure cell readiness.
2) Experiments fused cells at different phases and observed activation, identifying maturation promoting factor (MPF) as important for mitosis.
3) Further work identified MPF as a cyclin-Cdk complex, with cyclin levels and Cdk activating phosphorylations regulating progression through the cell cycle.
4) The cyclin B-Cdk1 complex promotes mitosis, driving events like nuclear envelope breakdown, before cyclin degradation inactivates Cdk1
Seminar delivered by Dr Gary Kerr at the University of Dundee on 13 Feb 2013 on meiotic nuclear divisions in budding yeast require PP2ACdc55-mediated antagonism of Net1 phosphorylation by Cdk
WDR7 up-regulation upon knocking down of neighboring noncoding RNA using siRN...Vahid Erfani-Moghadam
1) The document examines the effect of silencing the long non-coding RNA lincRNA-RoR using siRNAs delivered by polyamidoamine dendrimers on the expression of the neighboring WDR7 gene in breast cancer cells.
2) The study showed that lincRNA-RoR expression was significantly decreased after transfection with siRNA encapsulated in PAMAM dendrimers. WDR7 expression correspondingly increased significantly upon lincRNA-RoR silencing.
3) This study demonstrates that PAMAM dendrimers can effectively deliver siRNAs to silence lincRNA-RoR and upregulate the adjacent WDR7 gene, suggesting their potential as a non-viral gene therapy vector
Characterizing RNF20 and RNF40 in Class Switching of B CellsCathy Tie
Cathy Tie conducted a study to characterize the roles of RNF20 and RNF40 in class switching of B cells. She extracted mRNA from B cells and used PCR to amplify RNF20 and RNF40 cDNA, but initial primers were ineffective. Objectives included testing new primers and determining if overexpressing or depleting RNF20/RNF40 impacts class switching. Successful amplification of RNF20 and RNF40 would involve cloning the genes into a plasmid for bacterial expression and reintroducing them into B cells to observe effects on class switching. This may provide insights into DNA damage response pathways in antibody diversification and cancer.
The document contains multiple choice questions about biology topics such as cell structure, DNA, RNA, protein synthesis, cell cycle, and mitosis. Specifically, questions are asked about the genetic code, transcription, translation, phases of mitosis like prophase and metaphase, cell organelles like mitochondria and ribosomes, and cellular processes like protein synthesis and cell division.
This document describes a study using Denaturing Gradient Gel Electrophoresis (DGGE) to differentiate variants of the Infectious Salmon Anemia Virus (ISAv). DGGE allows detection of single point mutations by separating PCR-generated DNA fragments of the same length but different sequence. The study examines variants in segments 5 and 6 of the ISAv genome, which are important for pathogenicity. DGGE analysis of the variants showed distinct banding patterns, demonstrating the technique's ability to distinguish single nucleotide polymorphisms and its potential as a novel alternative for ISAv variant differentiation.
The document discusses induced pluripotent stem cells (iPSCs), which are adult cells that have been genetically reprogrammed to an embryonic stem cell-like state. Researchers found that forcing the expression of just four factors (Oct3/4, Sox2, Klf4, c-Myc) in mouse cells generated iPSCs that demonstrated pluripotency. Subsequently, similar reprogramming was achieved with human cells by several groups using varying combinations of reprogramming factors. iPSCs show potential for generating diverse cell types and even live organisms, offering opportunities for modeling diseases and developing regenerative therapies.
Kou Molecular and Cellular Biology 2003 3186-3201Jordan Irvin
This document summarizes a study analyzing mutations along the crystallographic dimer interface of the yeast TATA binding protein (TBP). 24 amino acids within the dimer interface were mutated. The mutants showed impaired dimerization, TAF1 binding, and TATA binding in vitro, consistent with structural models. In vivo, the mutants displayed phenotypes including low TBP levels, transcriptional derepression, slow growth, and synthetic toxicity with a TAF1 deletion, correlating with dimer instability. The results provide further support that TBP dimerization is physiologically important for negative gene regulation.
DNA damage repair Neil3 gene Knockout in MOLT-4iosrjce
RNAi is superannuated cellular mechanism that protect organism against viruses that replicate
through double- stranded RNA. RNAi can be used to diminish gene expression from plasmid expressing and
inserted sequence repeat. A stable harpin would be expressed after the vector was integrated into the genome.
In this paper a shiRNA expressing vector for Neil3 was designed and developed which is capable of replication
in MOLT-4. This shiRNA vector had the ability to arose the RNAi pathway, and reduce the gene expression of
Neil3. This was assessed by using pSilence 4.1CMV as a vector, and Gapdh as positive control.
TransVax, a therapeutic DNA vaccine targeting cytomegalovirus (CMV), showed promising results in a Phase 2 trial involving 80 hematopoietic cell transplant recipients. The vaccine significantly reduced CMV reactivation rates, increased the time to initial viral reactivation, and decreased the duration of viremia compared to placebo. No significant safety concerns were observed. The trial provides evidence that TransVax can control CMV reactivation in high-risk transplant patients in a manner superior to preemptive antiviral therapy alone.
Study of psymberin's mode of action using forward geneticsVincent Tsao
Through forward genetics screening in C. elegans, the researchers identified a mutation in the rpl-41 gene that confers resistance to psymberin, a potent cytotoxin. Using single nucleotide polymorphism (SNP) mapping, they narrowed the location of the mutation to a region between +3.1 cM and +3.5 cM on chromosome II. Sequencing of the mutant gene identified a point mutation that results in an amino acid change from proline to leucine. This mutation in the conserved rpl-41 gene, which encodes a 60S ribosomal protein, suggests psymberin's mechanism of inducing cell death may involve additional pathways beyond just translation inhibition.
The nucleotide sequence of a rat 18S rRNA gene was determined. The 18S rRNA encoded in the gene contains 1874 nucleotides. The sequences of rat and Xenopus laevis 18S rRNAs are very similar, with only minor differences like short insertions in the rat sequence. A secondary structure for rat 18S rRNA is proposed based on comparisons to 17 other small ribosomal subunit RNA sequences. While the primary sequences of rat and bacterial RNAs differ, their deduced secondary structures are remarkably similar.
This document summarizes key aspects of mantle cell lymphoma (MCL) biology and pathogenesis. It discusses:
1) MCL is characterized by the t(11;14) translocation resulting in cyclin D1 overexpression. Prognosis is poor with median survival of 3-4 years, though survival has improved with new therapies.
2) MCL pathogenesis involves dysregulation of cell cycle control and DNA damage response pathways. Genetic alterations affect genes like ATM, p53, ARF-INK4a locus, MDM2, CDK4/cyclin D, and RB.
3) MCL demonstrates a spectrum of variants from classical to blastoid based on proliferation. Bi
This study examined the effects of transgenic cotton expressing Bacillus thuringiensis (Bt) toxin on natural enemies of the cotton bollworm/legume pod borer, Helicoverpa armigera. The researchers found that Bt toxin was detected in cocoons of the parasitoid Campoletis chlorideae, but reduction in cocoon formation was due to early mortality of H. armigera rather than direct effects on the parasitoid. Feeding Bt-intoxicated H. armigera larvae to the predator Cheilomenes sexmaculatus increased its development time by 1.5 days but did not affect adult emergence. Bt-transgenic cotton-fed cotton a
Difference between prokaryotic and eukaryotic translationAman Ullah
This document discusses the differences between prokaryotic and eukaryotic translation but provides no other details as the link is broken and no text is available from the given source.
Biosafety of transgenic crops to natural enemies of Cotton Bollworm/Legume Po...ICRISAT
The study evaluated the effects of transgenic cotton expressing Bacillus thuringiensis (Bt) toxin on natural enemies of cotton bollworm/legume pod borer (Helicoverpa armigera). Field experiments showed egg parasitism was greater in Bt cotton, while cocoon formation of the larval parasitoid Campoletis chlorideae was lower due to early H. armigera mortality rather than direct Bt toxin effects. Laboratory experiments found Bt-intoxicated H. armigera increased development time of the predator Cheilomenes sexmaculatus by 1.5 days, and Bt cotton-fed aphids increased its development time up to 1.5 days
This document characterizes a diversity-generating retroelement (DGR) in Legionella pneumophila that confers adaptive advantages by diversifying a target protein. The DGR contains all the core components found in other DGR systems and is capable of transferring DNA sequence from a template repeat to a variable repeat of the target gene ldtA, accompanied by adenine-specific mutagenesis. The target protein LdtA is anchored in the bacterial outer membrane with its variable C-terminal domain exposed on the surface, allowing it to generate massive protein diversity on the bacterial surface. Analysis of related DGRs in other L. pneumophila strains showed they target different carrier proteins but maintain similar variable domains, demonstrating the modularity and adapt
A high copy number screen of the trs23Δ99C mutantGabriel Bendavit
This document summarizes a study investigating the role of the TRAPP I subunit TRS23, a guanine nucleotide exchange factor for the Ypt1/Rab GTPase, in the secretion pathway of yeast. A high copy number screen was performed on a yeast mutant with a C-terminal deletion of TRS23 (trs23Δ99C) to identify suppressors of its thermosensitive phenotype. From screening 36,000 transformed yeast colonies, 37 colonies showed suppression. Sequencing identified the gene CIN5, involved in chromosome instability, as a potential suppressor. Further experiments, such as isolating and transforming CIN5 into the trs23Δ99C mutant, are needed to confirm its role in suppress
This document discusses using lambda bacteriophage as a platform for phage display. It describes the major capsid proteins gpE and gpD, which forms trimers and decorates the capsid surface. Mutants called IPDF (inhibited by gpD-fusion) and supIPDF (suppressor of IPDF) were isolated that affect plating efficiency in the presence of gpD-fusions. Sequencing showed the mutations were not in gpD or gpE, but likely other capsid proteins like gpC that interact with them. Future work aims to generate mosaic lambda display particles by co-expressing gpD and gpD-fusions from modified lambda lacking gpD to study capsid incorporation.
This document reports the 1H, 13C, and 15N backbone resonance assignments of the 214 amino acid human DGCR8core protein (residues 493-706) determined using heteronuclear NMR spectroscopy. DGCR8core contains two tandem double-stranded RNA binding domains (dsRBDs) separated by a flexible linker that are required for recognizing and binding pri-miRNA substrates during miRNA biogenesis. The NMR assignments provide a foundation for further investigating the dynamics and RNA-binding properties of DGCR8core in solution. Secondary structure analysis using chemical shift indices matches the seven alpha helices and seven beta strands observed in the crystal structure of DGCR8core.
AJS: a lethal weapon to combat with cancers Treatment of Human Cancers Using...gan-navi
This document describes a lentiviral vector called AJS2001 that can selectively silence the Cdc6 gene in cancer cells. Cdc6 is essential for initiating DNA replication, and silencing it blocks cancer cell proliferation. The vector was shown to inhibit growth of several cancer cell lines through Cdc6 knockdown, inducing senescence. Case studies are presented of cancer patients who recovered after treatment with injections of AJS2001 lentivirus, including patients with gastric carcinoma, brain and liver metastases from ovarian cancer, lung and larynx metastases from parotid carcinoma, and breast cancer.
This document summarizes a thesis project investigating the regulation of the anaerobic responsive transcription factor TdcA in Salmonella Typhimurium. The project aims to construct Salmonella strains with deletions or tags of genes involved in regulating tdcA expression, and to analyze the effects on tdcA transcription and protein binding to its promoter region. Specifically, the project will create ∆tdcA and tdcA-FLAG strains to identify members of the TdcA regulon, and use transcriptional analysis and chromatin immunoprecipitation to study how proteins like CRP, FNR, and H-NS regulate tdcA expression in Salmonella. Understanding TdcA regulation is important because it plays a role in Salmon
The document summarizes key concepts about the cell cycle, including:
1) The cell cycle consists of interphase (G1, S, G2 phases) and mitosis (M phase), with checkpoints between phases to ensure cell readiness.
2) Experiments fused cells at different phases and observed activation, identifying maturation promoting factor (MPF) as important for mitosis.
3) Further work identified MPF as a cyclin-Cdk complex, with cyclin levels and Cdk activating phosphorylations regulating progression through the cell cycle.
4) The cyclin B-Cdk1 complex promotes mitosis, driving events like nuclear envelope breakdown, before cyclin degradation inactivates Cdk1
Seminar delivered by Dr Gary Kerr at the University of Dundee on 13 Feb 2013 on meiotic nuclear divisions in budding yeast require PP2ACdc55-mediated antagonism of Net1 phosphorylation by Cdk
WDR7 up-regulation upon knocking down of neighboring noncoding RNA using siRN...Vahid Erfani-Moghadam
1) The document examines the effect of silencing the long non-coding RNA lincRNA-RoR using siRNAs delivered by polyamidoamine dendrimers on the expression of the neighboring WDR7 gene in breast cancer cells.
2) The study showed that lincRNA-RoR expression was significantly decreased after transfection with siRNA encapsulated in PAMAM dendrimers. WDR7 expression correspondingly increased significantly upon lincRNA-RoR silencing.
3) This study demonstrates that PAMAM dendrimers can effectively deliver siRNAs to silence lincRNA-RoR and upregulate the adjacent WDR7 gene, suggesting their potential as a non-viral gene therapy vector
Characterizing RNF20 and RNF40 in Class Switching of B CellsCathy Tie
Cathy Tie conducted a study to characterize the roles of RNF20 and RNF40 in class switching of B cells. She extracted mRNA from B cells and used PCR to amplify RNF20 and RNF40 cDNA, but initial primers were ineffective. Objectives included testing new primers and determining if overexpressing or depleting RNF20/RNF40 impacts class switching. Successful amplification of RNF20 and RNF40 would involve cloning the genes into a plasmid for bacterial expression and reintroducing them into B cells to observe effects on class switching. This may provide insights into DNA damage response pathways in antibody diversification and cancer.
The document contains multiple choice questions about biology topics such as cell structure, DNA, RNA, protein synthesis, cell cycle, and mitosis. Specifically, questions are asked about the genetic code, transcription, translation, phases of mitosis like prophase and metaphase, cell organelles like mitochondria and ribosomes, and cellular processes like protein synthesis and cell division.
This document describes a study using Denaturing Gradient Gel Electrophoresis (DGGE) to differentiate variants of the Infectious Salmon Anemia Virus (ISAv). DGGE allows detection of single point mutations by separating PCR-generated DNA fragments of the same length but different sequence. The study examines variants in segments 5 and 6 of the ISAv genome, which are important for pathogenicity. DGGE analysis of the variants showed distinct banding patterns, demonstrating the technique's ability to distinguish single nucleotide polymorphisms and its potential as a novel alternative for ISAv variant differentiation.
The document discusses induced pluripotent stem cells (iPSCs), which are adult cells that have been genetically reprogrammed to an embryonic stem cell-like state. Researchers found that forcing the expression of just four factors (Oct3/4, Sox2, Klf4, c-Myc) in mouse cells generated iPSCs that demonstrated pluripotency. Subsequently, similar reprogramming was achieved with human cells by several groups using varying combinations of reprogramming factors. iPSCs show potential for generating diverse cell types and even live organisms, offering opportunities for modeling diseases and developing regenerative therapies.
Kou Molecular and Cellular Biology 2003 3186-3201Jordan Irvin
This document summarizes a study analyzing mutations along the crystallographic dimer interface of the yeast TATA binding protein (TBP). 24 amino acids within the dimer interface were mutated. The mutants showed impaired dimerization, TAF1 binding, and TATA binding in vitro, consistent with structural models. In vivo, the mutants displayed phenotypes including low TBP levels, transcriptional derepression, slow growth, and synthetic toxicity with a TAF1 deletion, correlating with dimer instability. The results provide further support that TBP dimerization is physiologically important for negative gene regulation.
DNA damage repair Neil3 gene Knockout in MOLT-4iosrjce
RNAi is superannuated cellular mechanism that protect organism against viruses that replicate
through double- stranded RNA. RNAi can be used to diminish gene expression from plasmid expressing and
inserted sequence repeat. A stable harpin would be expressed after the vector was integrated into the genome.
In this paper a shiRNA expressing vector for Neil3 was designed and developed which is capable of replication
in MOLT-4. This shiRNA vector had the ability to arose the RNAi pathway, and reduce the gene expression of
Neil3. This was assessed by using pSilence 4.1CMV as a vector, and Gapdh as positive control.
TransVax, a therapeutic DNA vaccine targeting cytomegalovirus (CMV), showed promising results in a Phase 2 trial involving 80 hematopoietic cell transplant recipients. The vaccine significantly reduced CMV reactivation rates, increased the time to initial viral reactivation, and decreased the duration of viremia compared to placebo. No significant safety concerns were observed. The trial provides evidence that TransVax can control CMV reactivation in high-risk transplant patients in a manner superior to preemptive antiviral therapy alone.
Study of psymberin's mode of action using forward geneticsVincent Tsao
Through forward genetics screening in C. elegans, the researchers identified a mutation in the rpl-41 gene that confers resistance to psymberin, a potent cytotoxin. Using single nucleotide polymorphism (SNP) mapping, they narrowed the location of the mutation to a region between +3.1 cM and +3.5 cM on chromosome II. Sequencing of the mutant gene identified a point mutation that results in an amino acid change from proline to leucine. This mutation in the conserved rpl-41 gene, which encodes a 60S ribosomal protein, suggests psymberin's mechanism of inducing cell death may involve additional pathways beyond just translation inhibition.
The nucleotide sequence of a rat 18S rRNA gene was determined. The 18S rRNA encoded in the gene contains 1874 nucleotides. The sequences of rat and Xenopus laevis 18S rRNAs are very similar, with only minor differences like short insertions in the rat sequence. A secondary structure for rat 18S rRNA is proposed based on comparisons to 17 other small ribosomal subunit RNA sequences. While the primary sequences of rat and bacterial RNAs differ, their deduced secondary structures are remarkably similar.
This document summarizes key aspects of mantle cell lymphoma (MCL) biology and pathogenesis. It discusses:
1) MCL is characterized by the t(11;14) translocation resulting in cyclin D1 overexpression. Prognosis is poor with median survival of 3-4 years, though survival has improved with new therapies.
2) MCL pathogenesis involves dysregulation of cell cycle control and DNA damage response pathways. Genetic alterations affect genes like ATM, p53, ARF-INK4a locus, MDM2, CDK4/cyclin D, and RB.
3) MCL demonstrates a spectrum of variants from classical to blastoid based on proliferation. Bi
This study examined the effects of transgenic cotton expressing Bacillus thuringiensis (Bt) toxin on natural enemies of the cotton bollworm/legume pod borer, Helicoverpa armigera. The researchers found that Bt toxin was detected in cocoons of the parasitoid Campoletis chlorideae, but reduction in cocoon formation was due to early mortality of H. armigera rather than direct effects on the parasitoid. Feeding Bt-intoxicated H. armigera larvae to the predator Cheilomenes sexmaculatus increased its development time by 1.5 days but did not affect adult emergence. Bt-transgenic cotton-fed cotton a
Difference between prokaryotic and eukaryotic translationAman Ullah
This document discusses the differences between prokaryotic and eukaryotic translation but provides no other details as the link is broken and no text is available from the given source.
Biosafety of transgenic crops to natural enemies of Cotton Bollworm/Legume Po...ICRISAT
The study evaluated the effects of transgenic cotton expressing Bacillus thuringiensis (Bt) toxin on natural enemies of cotton bollworm/legume pod borer (Helicoverpa armigera). Field experiments showed egg parasitism was greater in Bt cotton, while cocoon formation of the larval parasitoid Campoletis chlorideae was lower due to early H. armigera mortality rather than direct Bt toxin effects. Laboratory experiments found Bt-intoxicated H. armigera increased development time of the predator Cheilomenes sexmaculatus by 1.5 days, and Bt cotton-fed aphids increased its development time up to 1.5 days
Biosaftey issues related to gm crops and transgenic variety release Sachin Ekatpure
The document discusses issues related to biosafety and registration of transgenic agricultural organisms. It outlines three main biosafety concerns: environmental safety, food safety for human and animal health, and risk management. Some potential environmental risks discussed include effects on non-target organisms, development of insect resistance, gene flow, increased weediness, loss of biodiversity, changes in soil ecology, and genetic contamination. Food safety concerns include toxicity, allergenicity, and unintended effects. The document also describes India's biosafety regulatory framework and approval process for transgenic crops, which involves biosafety assessment and approval from multiple government committees and agencies before crops can be cultivated and marketed.
Translation in prokaryotes involves three main stages - initiation, elongation, and termination. During initiation, the small ribosomal subunit binds to an mRNA with help from initiation factors and forms the initiation complex. In elongation, tRNAs bring amino acids to the ribosome according to mRNA codons and peptide bonds are formed. Termination occurs when a stop codon signals for the release of the complete polypeptide from the ribosome. The process requires several RNA and protein molecules working together, including ribosomes, tRNAs, and various factors, to translate the genetic message into a polypeptide product.
Translation is the process by which proteins are synthesized from messenger RNA (mRNA) in eukaryotes, which are organisms with membrane-bound nuclei. Translation involves mRNA being decoded on ribosomes into a polypeptide chain. It occurs through three main steps - initiation, elongation, and termination. Initiation involves the small ribosomal subunit binding to the 5' end of mRNA and scanning for the start codon. Elongation is the sequential addition of amino acids specified by the mRNA codons. Termination occurs when a stop codon is reached and release factors cause the ribosome to dissociate and release the completed protein.
Phytoremediation is the process of using plants to remove contamination from soil or water. It involves using plants and their associated microorganisms in the rhizosphere to degrade, contain, or remove pollutants from the environment. Some key advantages are that it is a cost-effective, environmentally friendly way to remediate large areas of contaminated land. However, it is limited to sites with lower contaminant concentrations and works more slowly than conventional remediation methods. Common contaminants removed through phytoremediation include heavy metals, hydrocarbons, pesticides, and explosives. The process works through plants absorbing, degrading, or stabilizing pollutants in their tissues or the surrounding soil.
This document summarizes the process of translation. Translation is the process by which the sequence of nucleotides in messenger RNA directs the incorporation of amino acids into a protein. It involves three main steps - initiation, elongation, and termination. Initiation requires various initiation factors and ribosomal subunits to form the initiation complex. Elongation is a cyclic process of aminoacyl-tRNA binding, peptide bond formation, and translocation. Termination occurs when a stop codon is reached, releasing the polypeptide chain. Ribosomal recycling then dissociates the post-termination complex to prepare the ribosome for another round of translation.
Phytoremediation uses plants to remove contaminants from soil or water. It is a more environmentally friendly and cost-effective alternative to excavating and disposing of contaminated material elsewhere. Various phytoremediation processes include phytoextraction, which uses plants to remove contaminants from soil into harvestable biomass, phytostabilization which stabilizes contaminants in soil near roots, and phytotransformation which transforms contaminants via plant metabolism. Genetic engineering can enhance natural phytoremediation capabilities.
Apoptosis, also known as programmed cell death, is a natural process by which cells self-destruct in response to internal or external signals. It is distinct from necrosis in that it involves chromatin condensation, cell shrinkage, and preservation of organelles, allowing for rapid engulfment by neighboring cells without inflammation. Apoptosis is initiated through either the intrinsic mitochondrial pathway or the extrinsic death receptor pathway and is executed by caspases, a family of cysteine proteases. It plays an essential role in development and homeostasis by removing damaged or unneeded cells.
The document summarizes the central dogma of molecular biology and the processes of protein synthesis - transcription and translation. It explains that DNA is transcribed into mRNA in the nucleus, and mRNA is then translated into proteins by ribosomes in the cytoplasm. Transcription involves RNA polymerase copying the DNA code into mRNA. Translation involves tRNAs matching their anticodons to the mRNA codons and adding the corresponding amino acids to form a polypeptide chain. The genetic code is universal and degenerate, with multiple codons coding for each amino acid.
Transcription is the process of making mRNA from DNA. It involves RNA polymerase making an mRNA complementary copy of the DNA strand. Translation is the process of using the mRNA to produce a polypeptide by linking amino acids specified by the mRNA codons. During transcription, introns are removed from pre-mRNA and exons are joined to form mature mRNA. If the parent DNA strand is A A T G C A G T, the complementary mRNA strand will be U U A C G U C A.
The document summarizes the process of translation in cells. It describes how the genetic code on mRNA is translated into a sequence of amino acids with the help of tRNA and ribosomes. There are several steps involved: 1) amino acids are activated by attaching to tRNA, 2) initiation begins with formation of initiation complexes on ribosomes, 3) elongation occurs as new amino acids are added one by one to the growing polypeptide chain through movement of tRNA between ribosomal sites, and 4) termination releases the full protein when a stop codon is reached. The entire process requires energy in the form of GTP hydrolysis at several steps.
The document defines and describes various types of translation including:
- Oral and written translation which can be done consecutively or simultaneously
- Computer-assisted translation which uses computer programs to aid the human translation process
- Machine translation which uses computer programs to translate without human intervention
It also discusses different types of translation based on factors like the unit, aim, tasks/objectives, and number of translators involved. Some translation types discussed include: literal, idiomatic, committee, common language, dynamic, and thought-for-thought translations.
This document summarizes research characterizing the intracellular localization of the SNARE protein VTI13. Key points:
- VTI13 was found to localize to the vacuole membrane and early endosomes/trans-Golgi network in Arabidopsis root hair cells.
- The researchers cloned an mCherry-RABA1g fusion protein to use as an early endosome marker. They also constructed GFP-VTI13.
- GFP-VTI13 was found to localize to the same compartments (vacuole and punctate bodies) in tobacco leaf cells as in Arabidopsis, validating the experimental system.
- Preliminary co-localization experiments suggest GFP-V
Mir193b–365 is essential for brown fat differentiation by regulating genes involved in adipogenesis. The study identified Mir193b-365 as a microRNA complex necessary for brown adipose tissue differentiation. Blocking Mir193b expression inhibited brown fat marker genes, pointing to its critical role in brown fat development. Mir193b-365 associates closely with mRNAs like Prdm16 and Pparα that help upregulate it during differentiation, inducing adipogenic factors while suppressing myogenic factors.
Mdm2 oncoprotein. Cell Mol Life Sci 55:96–107
1) Researchers identified a novel oncogene, ribosomal protein L10a (RPL10a), using a cDNA library from Daudi cells to transform p53 knockout mouse embryonic fibroblast cells.
2) Western blot analysis of tumors from myc mice showed different levels of p53 and ARF expression, indicating deregulation of the myc-ARF-p53 pathway.
3) Sequencing of DNA from foci identified RPL10a as a potential oncogene, and its interaction with Mdm2 may imitate effects of myc overexpression, contributing to tumor progression in the absence of p53
Plasmids are small, autonomous DNA molecules that can replicate independently of the bacterial chromosome. They are useful as vectors in genetic engineering due to their ability to replicate autonomously and contain selectable marker genes. This document discusses the characteristics and functions of various natural and artificial plasmid vectors used in E. coli, including pSC101, pBR322, pUC series, and pGEMR. Key features of these vectors like size, copy number, restriction sites, resistance genes, and applications are described.
RAS is one of the most frequently mutated oncogenes in human cancer. KRAS is the isoform most frequently mutated, which constitutes about 85% of RAS mutations. As the most frequently mutated RAS isoform, KRAS is intensively studied in the past years.
In the formulation of KRAS integrated research plan, Medicilon has in-depth communication with customers. The backbone of scientific research has combined the characteristics of each case with years of practical experience and technical accumulation, and carefully submitted high-quality experimental plans and results to customers. Medicilon provides KRAS-targeted drug discovery, CMC research (API + formulation), pharmacodynamics research, PK study, safety evaluation and other services.https://www.medicilon.com/platform/kras/
This study examined 51 Brazilian Plasmodium falciparum isolates for polymorphisms in the Pfmdr1 gene thought to be associated with chloroquine resistance. 49 of the isolates were found to be resistant to chloroquine in vitro, while all were sensitive to mefloquine, amodiaquine, and quinine. The isolates were analyzed for three Pfmdr1 polymorphisms: Asn86Tyr, Asn1042Asp, and Asp1246Tyr. Asn86Tyr was not detected in any isolates, while Asn1042Asp was found in 50 isolates and Asp1246Tyr was found in all 51 isolates. This provides support that As
A CRISPR/Cas9, works like a biological version of a word-processing programme’s “find and replace”. Its simplicity and extremely low cost of implementation is the reason to use. How Cas 9 is activated and its mechanism (DNA binding and cleavage), it's regulation and application in human disease therapy, new drug screening, agriculture and biofuel etc.
The document summarizes the MAPK-P38 pathway, also known as the death pathway. It discusses that p38 was discovered in cells treated with inflammatory cytokines and as the target of an anti-inflammatory drug. It describes the four p38 subfamily members identified through cloning strategies and notes they all contain the TGY sequence in their activation loops. The document also outlines that p38 isoforms are differentially sensitive to inhibitors and can be activated by various stresses and stimuli. It discusses a splice variant of p38 called Mxi that has unique properties compared to p38a in terms of activation and insensitivity to inhibitors. In closing, it mentions the kinases MEK3 and MEK6 that activate p38 family members
By constructing a plasmid containing flanking sequences of the adenylate cyclase gene and a tryptophan marker gene, researchers aim to knockout the adenylate cyclase gene in Schizophyllum commune via homologous recombination. The plasmid would replace the adenylate cyclase sequence with the tryptophan gene in the genome, allowing only cells without adenylate cyclase to grow in media lacking tryptophan. Using a ku80 knockout strain of S. commune increases the likelihood of homologous recombination during transformation. Primers and restriction enzymes were used to amplify flanking sequences and insert them into a vector plasmid to ultimately construct the adenylate cyclase knockout plasmid.
Christine Williams reviews various technologies for detecting cyclic AMP (cAMP) levels in high-throughput screening assays. The review highlights radiometric, fluorescence polarization, luminescence, and electrochemiluminescence methods. It emphasizes practical considerations for choosing an assay to identify modulators of G protein-coupled receptors that signal through cAMP. Future technologies may provide even greater biological information for drug discovery.
This study identified protein arginine methyltransferase 6 (PRMT6) as a coactivator of nuclear factor-kappa B (NF-κB) through three main findings:
1. Transgenic mice overexpressing a fusion of PRMT6 and the estrogen receptor displayed increased levels of interleukin 6 (IL-6), an NF-κB target gene, upon tamoxifen treatment.
2. PRMT6 was found to directly interact with the NF-κB subunit RelA and enhance the transcriptional activity of an NF-κB reporter.
3. PRMT6 was recruited by RelA to selective NF-κB target promoters upon TNF-α stimulation and its overexpression led to increased nuclear accumulation of Rel
The document discusses the plasmid vector pBR322, which was constructed in 1977 and is one of the most commonly used cloning vectors. It describes the origins and components of pBR322, including two antibiotic resistance genes, the origin of replication, and restriction enzyme cleavage sites. The document also summarizes the construction of several derivatives of pBR322, including pBR327, pUC18, and pBR118/119, and notes their applications and advantages over the original pBR322 vector.
An aminopeptidase P (PepP)-encoding gene has been cloned from Streptomyces lividans 66. The gene, pepP, was localized by deletion mapping and its nucleotide sequence was determined. The deduced amino acid sequence was found to display significant similarity to Escherichia coli PepP. The partially purified S. lividans enzyme had a 50-kDa subunit and was present as a homodimer, confirming that pepP encodes the observed intracellular PepP.
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE B.pdfamzonknr
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE
BACKGROUND CONTEXT Lab: Differential Expression Differential gene expression provides
the ability for a cell or organism to respond to a constantly changing external environment. The
specific constellation of proteins required for optimal function and growth varies with cellular
age and environmental context. Thus, protein production is carefully regulated by multiple
mechanisms that modulate both transcriptional and translational pathways. Control of
transcription initiation by RNA polymerase is a predominant mechanism for regulating
expression of specific proteins, presumably because it provides maximal conservation of energy
for the cell. We can often observe the consequence of differential transcription due to the
presence or absence of particular proteins or the growth in particular environments. Control can
also occur at translation; the mRNA is synthesized, but only in certain circumstances is it
translated. Control can also occur at the level of protein function; the protein is inactive, or
activity is not observed due to the lack of the substrate. In this lab we will observe differential
expression of two different genes encoded on plasmids. We will analyze transcriptional activity,
translational activity, and protein function. Plasmids are extra-chromosomal DNA. Bacteria often
have plasmids and will replicate the plasmid and pass it to daughter cells (vertical transmission)
and to neighboring cells (horizontal). Plasmids are a mechanism of gene diversity. In order to
stably retain the plasmid, there needs to be some type of metabolic reason for the bacteria to
maintain the plasmid. In other words, the plasmid confers an advantage. Plasmids contain: 1. Ori:
the plasmid may present is low or high copy number. 2. Lab generated plasmids typically also
contain a selectable marker (antibiotic resistance), 3. Additional gene for ease of visual screening
4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant.
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE BAC.pdfamzonknr
ONLY THE LAST QUESTION IS THE POINT OF POST. THE OTHER PAGES ARE
BACKGROUND CONTEXT Lab: Differential Expression Differential gene expression provides
the ability for a cell or organism to respond to a constantly changing external environment. The
specific constellation of proteins required for optimal function and growth varies with cellular
age and environmental context. Thus, protein production is carefully regulated by multiple
mechanisms that modulate both transcriptional and translational pathways. Control of
transcription initiation by RNA polymerase is a predominant mechanism for regulating
expression of specific proteins, presumably because it provides maximal conservation of energy
for the cell. We can often observe the consequence of differential transcription due to the
presence or absence of particular proteins or the growth in particular environments. Control can
also occur at translation; the mRNA is synthesized, but only in certain circumstances is it
translated. Control can also occur at the level of protein function; the protein is inactive, or
activity is not observed due to the lack of the substrate. In this lab we will observe differential
expression of two different genes encoded on plasmids. We will analyze transcriptional activity,
translational activity, and protein function. Plasmids are extra-chromosomal DNA. Bacteria often
have plasmids and will replicate the plasmid and pass it to daughter cells (vertical transmission)
and to neighboring cells (horizontal). Plasmids are a mechanism of gene diversity. In order to
stably retain the plasmid, there needs to be some type of metabolic reason for the bacteria to
maintain the plasmid. In other words, the plasmid confers an advantage. Plasmids contain: 1. Ori:
the plasmid may present is low or high copy number. 2. Lab generated plasmids typically also
contain a selectable marker (antibiotic resistance), 3. Additional gene for ease of visual screening
4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant.
Polymerase chain reaction (PCR) is a technique used to amplify DNA sequences. It has numerous applications including prenatal diagnosis, forensics, detecting viruses like HIV, and cancer research. PCR involves denaturing DNA, annealing primers, and extending new strands through multiple cycles. Real-time PCR uses probes to detect amplification during each cycle, allowing quantification. It has revolutionized fields like molecular biology and genetics through rapid copying of DNA.
This document summarizes research into the mechanisms that determine which strand of an RNA duplex becomes the guide strand that is incorporated into the RNA-induced silencing complex (RISC) during microRNA processing in mammals. Through experiments manipulating the 5' nucleotides and thermodynamic stability of engineered miRNA precursors, the researchers found that guide strand selection is governed by two independent "rules": the 5'-nucleotide selection rule favors strands starting with U or A, while the thermodynamic stability rule favors the less stably base-paired end. They propose a "twin drive" model where the MID domain of Argonaute proteins sense both rules simultaneously through recognition of 5' nucleotides and phosphates.
Lab: Differential Expression Differential gene expression provides the ability for a cell or
organism to respond to a constantly changing external environment. The specific constellation of
proteins required for optimal function and growth varies with cellular age and environmental
context. Thus, protein production is carefully regulated by multiple mechanisms that modulate
both transcriptional and translational pathways. Control of transcription initiation by RNA
polymerase is a predominant mechanism for regulating expression of specific proteins,
presumably because it provides maximal conservation of energy for the cell. We can often
observe the consequence of differential transcription due to the presence or absence of particular
proteins or the growth in particular environments. Control can also occur at translation; the
mRNA is synthesized, but only in certain circumstances is it translated. Control can also occur at
the level of protein function; the protein is inactive, or activity is not observed due to the lack of
the substrate. In this lab we will observe differential expression of two different genes encoded
on plasmids. We will analyze transcriptional activity, translational activity, and protein function.
Plasmids are extra-chromosomal DNA. Bacteria often have plasmids and will replicate the
plasmid and pass it to daughter cells (vertical transmission) and to neighboring cells (horizontal).
Plasmids are a mechanism of gene diversity. In order to stably retain the plasmid, there needs to
be some type of metabolic reason for the bacteria to maintain the plasmid. In other words, the
plasmid confers an advantage. Plasmids contain: 1. Ori: the plasmid may present is low or high
copy number. 2. Lab generated plasmids typically also contain a selectable marker (antibiotic
resistance), 3. Additional gene for ease of visual screening 4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant plasmids. The transformed cells containing the plasmid with the gene of interest ca.
Lab: Differential Expression Differential gene expression provides the ability for a cell or
organism to respond to a constantly changing external environment. The specific constellation of
proteins required for optimal function and growth varies with cellular age and environmental
context. Thus, protein production is carefully regulated by multiple mechanisms that modulate
both transcriptional and translational pathways. Control of transcription initiation by RNA
polymerase is a predominant mechanism for regulating expression of specific proteins,
presumably because it provides maximal conservation of energy for the cell. We can often
observe the consequence of differential transcription due to the presence or absence of particular
proteins or the growth in particular environments. Control can also occur at translation; the
mRNA is synthesized, but only in certain circumstances is it translated. Control can also occur at
the level of protein function; the protein is inactive, or activity is not observed due to the lack of
the substrate. In this lab we will observe differential expression of two different genes encoded
on plasmids. We will analyze transcriptional activity, translational activity, and protein function.
Plasmids are extra-chromosomal DNA. Bacteria often have plasmids and will replicate the
plasmid and pass it to daughter cells (vertical transmission) and to neighboring cells (horizontal).
Plasmids are a mechanism of gene diversity. In order to stably retain the plasmid, there needs to
be some type of metabolic reason for the bacteria to maintain the plasmid. In other words, the
plasmid confers an advantage. Plasmids contain: 1. Ori: the plasmid may present is low or high
copy number. 2. Lab generated plasmids typically also contain a selectable marker (antibiotic
resistance), 3. Additional gene for ease of visual screening 4. Multiple cloning site
pUC19 is one of a series of plasmid cloning vectors created by Joachim Messing and co-workers.
The designation "pUC" is derived from the classical "p" prefix (denoting "plasmid") and the
abbreviation for the University of California, where early work on the plasmid series had been
conducted. It is a circular double stranded DNA and has 2686 base pairs. pUC19 is one of the
most widely used vector molecules as the recombinants, or the cells into which foreign DNA has
been introduced, can be easily distinguished from the non-recombinants based on color
differences of colonies on growth media. pUC18 is similar to pUC19, but the MCS region is
reversed. - pUC 19 has an origin of replication and is maintained at a high copy number. -
pUC19 encodes for an ampicillin resistance gene (amopR), via a -lactamase enzyme that
functions by degrading ampicillin and reducing its toxicity to the host. - It has an N-terminal
fragment of -galactosidase (lacZ) gene of E. coli which allows for visual screening of
recombinant plasmids. The transformed cells containing the plasmid with the gene of interest ca.
Similar to Analysis of mutant parasites that have a defect in adhésion (20)
Guide pour le suivi et l'évaluation des programmes - Séances Pratiques de la 5e édition du Cours international « Atelier Paludisme » - Luciano TUSEO - World Health Organization / Roll Back Malaria - Office for Madagascar and Reunion - Antananarivo, Madagascar - maloms@iris.mg
Monitoring and Evaluation Toolkit - Séances Pratiques de la 5e édition du Cours international « Atelier Paludisme » - Luciano TUSEO - World Health Organization / Roll Back Malaria - Office for Madagascar and Reunion - Antananarivo, Madagascar - maloms@iris.mg
Développement nouveaux médicaments - Séances Pratiques de la 5e édition du Cours international « Atelier Paludisme » - Pascal MILLET - Université Victor Segalen Bordeaux2, France - pascal.millet@u-bordeaux2.fr
Diagnostic biologique du paludisme - Séances Pratiques de la 5e édition du Cours international « Atelier Paludisme » - Didier MENARD et Vincent THONIER
Gene therapy can be broadly defined as the transfer of genetic material to cure a disease or at least to improve the clinical status of a patient.
One of the basic concepts of gene therapy is to transform viruses into genetic shuttles, which will deliver the gene of interest into the target cells.
Safe methods have been devised to do this, using several viral and non-viral vectors.
In the future, this technique may allow doctors to treat a disorder by inserting a gene into a patient's cells instead of using drugs or surgery.
The biggest hurdle faced by medical research in gene therapy is the availability of effective gene-carrying vectors that meet all of the following criteria:
Protection of transgene or genetic cargo from degradative action of systemic and endonucleases,
Delivery of genetic material to the target site, i.e., either cell cytoplasm or nucleus,
Low potential of triggering unwanted immune responses or genotoxicity,
Economical and feasible availability for patients .
Viruses are naturally evolved vehicles that efficiently transfer their genes into host cells.
Choice of viral vector is dependent on gene transfer efficiency, capacity to carry foreign genes, toxicity, stability, immune responses towards viral antigens and potential viral recombination.
There are a wide variety of vectors used to deliver DNA or oligo nucleotides into mammalian cells, either in vitro or in vivo.
The most common vector system based on retroviruses, adenoviruses, herpes simplex viruses, adeno associated viruses.
Computer in pharmaceutical research and development-Mpharm(Pharmaceutics)MuskanShingari
Statistics- Statistics is the science of collecting, organizing, presenting, analyzing and interpreting numerical data to assist in making more effective decisions.
A statistics is a measure which is used to estimate the population parameter
Parameters-It is used to describe the properties of an entire population.
Examples-Measures of central tendency Dispersion, Variance, Standard Deviation (SD), Absolute Error, Mean Absolute Error (MAE), Eigen Value
Nutritional deficiency Disorder are problems in india.
It is very important to learn about Indian child's nutritional parameters as well the Disease related to alteration in their Nutrition.
Pictorial and detailed description of patellar instability with sign and symptoms and how to diagnose , what investigations you should go with and how to approach with treatment options . I have presented this slide in my 2nd year junior residency in orthopedics at LLRM medical college Meerut and got good reviews for it
After getting it read you will definitely understand the topic.
- Video recording of this lecture in English language: https://youtu.be/Pt1nA32sdHQ
- Video recording of this lecture in Arabic language: https://youtu.be/uFdc9F0rlP0
- Link to download the book free: https://nephrotube.blogspot.com/p/nephrotube-nephrology-books.html
- Link to NephroTube website: www.NephroTube.com
- Link to NephroTube social media accounts: https://nephrotube.blogspot.com/p/join-nephrotube-on-social-media.html
TEST BANK For Brunner and Suddarth's Textbook of Medical-Surgical Nursing, 14...Donc Test
TEST BANK For Brunner and Suddarth's Textbook of Medical-Surgical Nursing, 14th Edition (Hinkle, 2017) Verified Chapter's 1 - 73 Complete.pdf
TEST BANK For Brunner and Suddarth's Textbook of Medical-Surgical Nursing, 14th Edition (Hinkle, 2017) Verified Chapter's 1 - 73 Complete.pdf
TEST BANK For Brunner and Suddarth's Textbook of Medical-Surgical Nursing, 14th Edition (Hinkle, 2017) Verified Chapter's 1 - 73 Complete.pdf
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdfOsvaldo Bernardo Muchanga
GASTROINTESTINAL INFECTIONS AND GASTRITIS
Osvaldo Bernardo Muchanga
Gastrointestinal Infections
GASTROINTESTINAL INFECTIONS result from the ingestion of pathogens that cause infections at the level of this tract, generally being transmitted by food, water and hands contaminated by microorganisms such as E. coli, Salmonella, Shigella, Vibrio cholerae, Campylobacter, Staphylococcus, Rotavirus among others that are generally contained in feces, thus configuring a FECAL-ORAL type of transmission.
Among the factors that lead to the occurrence of gastrointestinal infections are the hygienic and sanitary deficiencies that characterize our markets and other places where raw or cooked food is sold, poor environmental sanitation in communities, deficiencies in water treatment (or in the process of its plumbing), risky hygienic-sanitary habits (not washing hands after major and/or minor needs), among others.
These are generally consequences (signs and symptoms) resulting from gastrointestinal infections: diarrhea, vomiting, fever and malaise, among others.
The treatment consists of replacing lost liquids and electrolytes (drinking drinking water and other recommended liquids, including consumption of juicy fruits such as papayas, apples, pears, among others that contain water in their composition).
To prevent this, it is necessary to promote health education, improve the hygienic-sanitary conditions of markets and communities in general as a way of promoting, preserving and prolonging PUBLIC HEALTH.
Gastritis and Gastric Health
Gastric Health is one of the most relevant concerns in human health, with gastrointestinal infections being among the main illnesses that affect humans.
Among gastric problems, we have GASTRITIS AND GASTRIC ULCERS as the main public health problems. Gastritis and gastric ulcers normally result from inflammation and corrosion of the walls of the stomach (gastric mucosa) and are generally associated (caused) by the bacterium Helicobacter pylor, which, according to the literature, this bacterium settles on these walls (of the stomach) and starts to release urease that ends up altering the normal pH of the stomach (acid), which leads to inflammation and corrosion of the mucous membranes and consequent gastritis or ulcers, respectively.
In addition to bacterial infections, gastritis and gastric ulcers are associated with several factors, with emphasis on prolonged fasting, chemical substances including drugs, alcohol, foods with strong seasonings including chilli, which ends up causing inflammation of the stomach walls and/or corrosion. of the same, resulting in the appearance of wounds and consequent gastritis or ulcers, respectively.
Among patients with gastritis and/or ulcers, one of the dilemmas is associated with the foods to consume in order to minimize the sensation of pain and discomfort.
Breast cancer: Post menopausal endocrine therapyDr. Sumit KUMAR
Breast cancer in postmenopausal women with hormone receptor-positive (HR+) status is a common and complex condition that necessitates a multifaceted approach to management. HR+ breast cancer means that the cancer cells grow in response to hormones such as estrogen and progesterone. This subtype is prevalent among postmenopausal women and typically exhibits a more indolent course compared to other forms of breast cancer, which allows for a variety of treatment options.
Diagnosis and Staging
The diagnosis of HR+ breast cancer begins with clinical evaluation, imaging, and biopsy. Imaging modalities such as mammography, ultrasound, and MRI help in assessing the extent of the disease. Histopathological examination and immunohistochemical staining of the biopsy sample confirm the diagnosis and hormone receptor status by identifying the presence of estrogen receptors (ER) and progesterone receptors (PR) on the tumor cells.
Staging involves determining the size of the tumor (T), the involvement of regional lymph nodes (N), and the presence of distant metastasis (M). The American Joint Committee on Cancer (AJCC) staging system is commonly used. Accurate staging is critical as it guides treatment decisions.
Treatment Options
Endocrine Therapy
Endocrine therapy is the cornerstone of treatment for HR+ breast cancer in postmenopausal women. The primary goal is to reduce the levels of estrogen or block its effects on cancer cells. Commonly used agents include:
Selective Estrogen Receptor Modulators (SERMs): Tamoxifen is a SERM that binds to estrogen receptors, blocking estrogen from stimulating breast cancer cells. It is effective but may have side effects such as increased risk of endometrial cancer and thromboembolic events.
Aromatase Inhibitors (AIs): These drugs, including anastrozole, letrozole, and exemestane, lower estrogen levels by inhibiting the aromatase enzyme, which converts androgens to estrogen in peripheral tissues. AIs are generally preferred in postmenopausal women due to their efficacy and safety profile compared to tamoxifen.
Selective Estrogen Receptor Downregulators (SERDs): Fulvestrant is a SERD that degrades estrogen receptors and is used in cases where resistance to other endocrine therapies develops.
Combination Therapies
Combining endocrine therapy with other treatments enhances efficacy. Examples include:
Endocrine Therapy with CDK4/6 Inhibitors: Palbociclib, ribociclib, and abemaciclib are CDK4/6 inhibitors that, when combined with endocrine therapy, significantly improve progression-free survival in advanced HR+ breast cancer.
Endocrine Therapy with mTOR Inhibitors: Everolimus, an mTOR inhibitor, can be added to endocrine therapy for patients who have developed resistance to aromatase inhibitors.
Chemotherapy
Chemotherapy is generally reserved for patients with high-risk features, such as large tumor size, high-grade histology, or extensive lymph node involvement. Regimens often include anthracyclines and taxanes.
Tele Optometry (kunj'sppt) / Basics of tele optometry.
Analysis of mutant parasites that have a defect in adhésion
1. ANALYSIS OF MUTANT PARASITES
THAT HAVE A DEFECT IN ADHESION
Denise MATTEI
Cours International « Atelier Paludisme »
10 Mars au 18 Avril 1008 – Institut Pasteur de Madagascar
Mar08
2. TRAFFICKING OF VIRULENCE FACTORS IN INFECTED
RED BLOOD CELLS
SEARCH FOR TRAFFICKING
MUTANT PARASITES
Mar08
3. other
PfEMP1
molecules?
Band 3 rifin
Glycophorin C
Ankyrin
Adductin 4.1
Spectrin
4.2 4.9
Tropomyosin
and Actin
Tropomodulin
Knob-associated proteins
PfHRP-1
PfEMP-2
PfEMP-3
knobs
Mar08
4. OBSERVATIONS
CLAG = cytoadherence linked asexual gene
Identified through the study of parasite lines that no longer bound to C32
melanoma cells = subtelomeric deletion in chr 9 (Biggs, BA et al. PNAS,1989)
.
Genetic knock-out (Trenholme, KR et al. PNAS, 2000)
or
antisense RNA inhibition Gardiner, DL et al. MBP, 2000)
=> loss of the CD36 binding phenotype
.
CLAG9 may be involved in cytoadhesion by binding directly to CD36
or indirectly, through a role in PfEMP1 transport
Mar08
5. WORKING HYPOTHESIS
Is CLAG9 another CD36 binding molecule?
EXPERIMENTAL APPROACH
• P. falciparum lines selected to bind to CD36 or to CSA. Adhesion to
these receptors occurs in a mutually exclusive manner
Receptor «panning »
⇒ possibility to study the role of CLAG9 in
cytoadherent parasites that do not bind to CD36
CHO
CSA CD36
monomorphic parasites
Panned
FCR3
FCR3 CSA FCR3 CD36
• Specific antibodies Scherf et al., 1998, EMBO J. 17, 5418
=> to analyse the location of clag9 during the asexual blood stage development
Mar08
6. Transcription of the clag9 gene
The clag9 gene is transcribed in the
schizont stages of both CD36 and
CSA-selected parasites
3D7
- 9.49
- 7.46
- 4.40
hrs 2 6 10 14 18 22 26 30 34 38 42 46
Mar08
7. Clag9 is part of the RhopH complex in the rhoptries of
merozoites
r anti-clag9 mab7H8/50 nuclei merged
FITC TRICT DAPI
and
is transferred into the ring-stage parasite
r anti-clag9 mAb 4E10 merge DAPI
TRICT Oregon
green
Ling et al 2004. Mol.Microbiol.52:107
Mar08
8. Localisation of CLAG9 to the rhoptries
Clag9 RhopH2
Ling et al.2004. Mol.Microbiol.
Mar08
9. CLAG9
• clag9 gene is transcribed and expressed in late asexual
blood stages in CD36 and non-CD36 binders
• Clag9 is present in merozoites as part of the RhopH
complex in the rhoptries
• Anti-CLAG9 antibodies do not detect it at the
erythrocyte membrane of parasitized RBC
(trophozoites/schizonts)
Mar08
11. T9-96 and D10 do not cytoadhere to any known
adhesion receptor
FCR3CD36 D10
C32 cells
CD36 >> ICAM-1 >>CSA
Mar08
12. OBSERVATIONS
• T9-96 and D10 do not cytoadhere
• The complex RhopH is detected in the ring stages
HYPOTHESIS
clag9 is necessary to modify the parasite’s and/or
parasitophorous vacuole membrane
EXPERIMENTAL APPROACH
Analysis by immunofluorescence of clag9neg lines
and FCR3CD36
Mar08
13. RESULTS
The images were similar independently of the genotype
FCR3CD36
D10
T9.96
PfEMP1 Pf332 DAPI merge
Alexa TRICT
‘PfEMP1’ is located in association with the Maurer’s clefts
Mar08
14. FCR3CD36
D10
T9.96
Mab89 DAPI merge
Alexa
CONCLUSION
clag9neg parasites can export polypeptides
to the different cellular compartments
Mar08
15. OBSERVATIONS
• ‘ PfEMP1’-like molecule is exported in association with Maurer’s clefts
• ‘PfEMP1’ ?
• Is it exposed on the red blood cell surface ?
EXPERIMENTAL APPROACH
• Labelling of red blood cell surface of clag9neg strains
• Trypsin resistance
Mar08
16. CLAG9neg lines express a PfEMP1
surface protein
3 CD36
FCR
6
B
D10
A 100 µg Trypsin
T9.9
0 10
•High molecular weight
PfEMP1 •TX100-insol/SDS soluble
PfEMP1
•Trypsin sensitive
- 200kDa -
T9.96
Mar08
17. OBSERVATIONS
• The isolates T9.96 and D10 express PfEMP1 on the surface but do not
cytoadhere to any known receptor
HYPOTHESIS
• Adhesion negative parasites express a FUNCTIONAL var gene having
unique adhesive properties BUT can not switch expression to classical
var genes
• Adhesion negative parasites display a NON FUNCTIONAL conformation
* Post-translational modification to establish a functional state
(protease, kinase, etc…)
EXPERIMENTAL APPROACH
• Northern blot analysis
• Cloning and expression of the CIDR domain BINDING ASSAYS
Mar08
18. PfEMP1 is transcribed in the clag9neg
parasite lines T9.96 and D10
D10 FCR3CD36 D10 FCR3CD36
R S R S T Kb
R S R S T
8Kb - 9.49 -
- 7.46 -
- 4.40 -
- 2.57 -
- 1.35 -
D10cl9 ATS
Mar08
20. A classical var gene is transcribed and expressed
in mutant parasites
Semi-conserved Tandem
head structure association
NTS DBL1α
α CIDR1α
α DBL2δ
δ CIDR2β
β TM ATS
CR1 CD36 CD31
blood gr A CD31
heparin IgM
hep sulfate
ATS: acidic terminal segment; CIDR (α,β,γ): cisteine-rich interdomain region; DBL (α,β,γ,δ,ε): Duffy-binding-like domain;
NTS: N-terminal segment; TM: transmembrane domain;
Mar08
21. The CIDR domain from D10 is predicted to bind to CD36
M1 M2 M3
C9 C12
D10cl9 C L K NNKKTCGKKKCNRDCKCYE RW VKRKKEEFKKIKDHFGKQKD MQPYID PDMTLKILLNYVFLQ DMKDANG NPQHIAKIQELLE KKKVELEDNLNKNTIIDYMFEDDLEEINKC
Robinson et al. 2003.Mol.Microbiol. 47:1265-78
al. 2003.Mol.Microbiol. 47:1265-
Mar08
23. Adhesion on different purified receptors
FCR3 CD36 on differents receptors
CD36 ICAM qC1QR CSA CSC
D10 on differents receptors
CD36 ICAM qC1QR CSA CSC
Mar08
24. Binding assays with Dynal beads
Anti-CD36 labelled beads
CD36 protein
M2 domain on the surface
CIDR domain (M2 minimum CD36 binding site)-
pDisplay vector transfected in COS-7 cells
Mar08
25. CD36 Binding Properties of D10 CIDR1
Transfected anti-tag
COS7 cells merge
HA
D10
MC
+ control
CSA
(-) control
Mar08
26. Conclusions
• Cytoadhesion mutants express a PfEMP1 molecule at
the surface of the parasitized red blood cell
• The mutant parasites can not bind to the common
adhesion phenotypes (CD36, ICAM-1, CSA, Rosetting, etc.)
• Panning on endothelial cells does not restore adhesion
phenotype
Non-functional PfEMP1 surface expression !
M
35. HYPOTHESIS
• Adhesion negative parasites display a NON FUNCTIONAL conformation
* Post-translational modification to establish a functional state
(protease, kinase, etc…)
PERSPECTIVES
* Clag 9 KO => clag9 functional role
* other KO => chr9 region => genotype and phenotype analysis
* SYSTEMS BIOLOGY => RNAs transcribed/translated
ptn-ptn interactions cellular location
Mar08
36. National Institute for
BIHP (Institut Pasteur, Paris) Medical Research
(Mill Hill, London, UK)
Artur Scherf Tony Holder
Suwanna Chaorattanakawe Irene T. Ling
José Manuel L. Nogueira
Caroline Davis
Ehime University
Christine Scheidig-Benatar
Yvon Sterkers School of Medicine
(Ehime, Japan)
Emeric Roux Osamu Kaneko
Parasitologie Expérimentale
(Université de la Méditerranée, Marseille)
Cellular Microbiology and
Jürg Gysin
Catherine Lepolard Infectious Pathogeny
(Institute Pasteur Lille)
Frank Lafont
Sebastien Janel
37. ANALYSIS OF MUTANT PARASITES
THAT HAVE A DEFECT IN ADHESION
Denise MATTEI
Cours International « Atelier Paludisme »
10 Mars au 18 Avril 1008 – Institut Pasteur de Madagascar
Mar08
40. Why the PfEMP1expressed by the ‘clagneg’
isolates is non functional ?
HYPOTHESIS
• wrong conformation ?
• post-translational modification ?
how can it be tested?
apr07
41. Perspectives
• A NEW var GENE HAVING UNIQUE ASPECTS?
(non CD36, non CSA binding)
AND/OR
• A NEW HOST ENDOTHELIAL ADHESION RECEPTOR ?
OR
•A KNOWN PfEMP1 WHICH DISPLAYS A NON FUNCTIONAL
CONFORMATION
• clag9 KNOCK-OUT
42.
43. Analysis of surface-exposed PfEMP1 on PRBC
by trypsin cleavage
N N
R
FCR3 CD36 R
D10 kDa
B B
C C
Trypsin g 1000 100 0 1000 100 0
175
83
ATS 62
HRP
HRP
HSP 70 47.5
HSP 70
32.5
44. Analysis of surface-exposed PfEMP1 on PRBC
by trypsin and chymotrypsin cleavage
kDa D10 N FCR3 CD36 N
R R
B B
Trypsin C C
Chymotrypsin g 0 100 100 0 100 100
175
83
62 ATS
HRP
HRP
47.5 HSP 70
HSP 70
32.5
45. Identification of natural clag9 mutant
parasite lines
A 36
CS CD
6 C 7 6 3 3
9-9 D7 NRB 3D T9-9 FCR FCR
T 3 kDa
T9-96 3D7
r anti-clag9
175
clag9
chr9
83
62
r PI
47.5
32.5
clag9 probe
Mar08
46. Localisation of CLAG9
Clag9-GST 100 aa
pep1 pep2
r anti-clag9 mab7H8/50 nuclei merged
FITC TRICT DAPI
Clag9 is encoded by members of the clag multigene family
Ling,I et al. 2004. Mol.Microbiol. 52:107-118
Mar08
48. Perspectives
• A NEW var GENE HAVING UNIQUE ASPECTS?
(non CD36, non CSA binding)
AND/OR
• A NEW HOST ENDOTHELIAL ADHESION RECEPTOR ?
OR
• A KNOWN PfEMP1 WHICH DISPLAYS A
NON FUNCTIONAL CONFORMATION
• clag9 KNOCK-OUT
Mar08
49. Why PfEMP1 expressed by the ‘clag9neg’
isolates are non functional ?
HYPOTHESIS
* wrong conformation
* post-translational modification
Mar08