This document summarizes a study that investigated the timing of Xer recombination at dif sites during chromosome segregation in E. coli. The researchers developed a new quantitative PCR-based assay to directly measure recombination between dif sites in real time. Using this assay, they found that dif recombination only occurs shortly before or during cell division, even though the FtsK protein is recruited earlier to the septum. This suggests that FtsK activity is delayed relative to its recruitment. The precise coordination of FtsK function and septum assembly helps ensure that recombination only takes place once chromosomes have properly segregated.
This document summarizes information about bacteriophage vectors. It discusses how bacteriophages infect and replicate within bacterial cells, and how this process can be utilized to clone DNA fragments. Specifically, it describes commonly used phage vectors like lambda phage, M13 phage, and P1 phage. Lambda phage vectors can be insertion or replacement vectors, and allow cloning of fragments up to 24kb. M13 phage is a filamentous phage that can deliver single-stranded genomes up to 6.4kb. P1 phage can clone very large fragments up to 1000kb. In conclusion, bacteriophage vectors are useful tools for cloning large DNA fragments.
The document summarizes various genetic techniques including PCR, restriction mapping, the human genome project, in situ hybridization, and cloning the gene responsible for alkaptonuria. It provides an example of how PCR, genomic libraries, DNA sequencing, and other methods were used to clone the HGO gene involved in alkaptonuria. Ethical considerations are discussed around using genetic testing to predict late-onset genetic disorders in fetuses.
Wiki glossary epigenetic control of gene expressionBárbara Pérez
This document is a glossary defining terms related to epigenetics and gene expression. It defines over 80 key terms concisely, including definitions for adenosine triphosphate, alleles, autosomes, Barr body, bisulfite sequencing, blastocyst, boundary element, central dogma, chromatin, chromosome, cytosine methylation, DNA methylation, epigenetic reprogramming, epigenetics, embryonic stem cells, euchromatin, gene, genomic imprinting, histones, homologous chromosomes, imprinted genes, meiosis, mitosis, mRNA, nucleosome, placenta, and pluripotent cells. The glossary provides succinct yet informative definitions for fundamental concepts in molecular biology
This document summarizes recent evidence that the arrangement of chromosomes, gene loci, and nuclear bodies within the cell nucleus is not random but rather exhibits spatial organization that influences gene expression and nuclear processes. Techniques such as fluorescence in situ hybridization and chromosome conformation capture have provided insights into the positioning of chromosomes and genes relative to nuclear landmarks. Chromosomes occupy distinct territories within the nucleus and preferentially localize near the nuclear interior or periphery depending on their gene content. Association with nuclear subcompartments such as the nuclear lamina, nuclear pores, nucleoli, and polycomb bodies can impact the transcriptional state of genes and chromatin domains. Advances in genome-wide and time-lapse imaging approaches are helping to further characterize nuclear organization
This document discusses cloning vectors and plasmid-based cloning vectors. It provides information on plasmids, their structure, replication, and how they are used as cloning vectors. Some key points:
- Plasmids are small DNA molecules that can replicate independently of the bacterial chromosome and are often used as cloning vectors. They exist naturally in bacteria.
- Plasmids can have different structures depending on whether their DNA forms covalently closed circles or open circles. Supercoiled plasmids separate from open circles during electrophoresis.
- Plasmids can replicate in bacteria and confer phenotypes to their host like antibiotic resistance. They are categorized based on properties like being conjugative or copy number.
-
SYNTHETIC CHROMOSOME PLATFORMs IN PLANTS: CONCEPTS & APPLICATIONskundan Jadhao
This document outlines a seminar on synthetic/artificial chromosomes. It begins by discussing the need for synthetic chromosomes due to limitations of traditional genetic engineering approaches. It then describes various methods that have been used to develop artificial chromosomes, including the bottom-up and top-down methods. Several case studies are presented where telomere-mediated truncation was used to produce engineered minichromosomes in plants with endogenous centromeres. The document concludes by discussing ways that engineered minichromosomes can be amended in planta, such as through site-specific recombination systems and zinc finger nucleases.
This document discusses P1 vectors, which are cloning vectors derived from the P1 bacteriophage. P1 can transfer DNA between bacterial cells through transduction. The document describes:
1) The construction of P1 vectors pNS358 and pNS582, which contain P1 packaging and replication elements to clone and propagate large DNA fragments.
2) How the P1 system was used to clone DNA fragments up to 100 kb by packaging ligated vector/insert concatemers into phage heads.
3) Applications of P1 vectors including the creation of P1-derived artificial chromosomes (PACs) which can accommodate relatively large DNA fragments for cloning and mapping genomes.
Dna content,c value paradox, euchromatin heterochromatin, banding patternArchanaSoni3
DNA content refers to the amount of DNA in an organism's haploid chromosomes. It varies greatly between organisms, with eukaryotes generally having more DNA than prokaryotes. The amount of DNA does not always correlate with an organism's complexity, known as the C-value paradox. This is because eukaryotic DNA contains large amounts of non-coding repetitive sequences. Chromatin exists in two forms - euchromatin, which is less condensed and permits gene expression, and heterochromatin, which is highly condensed and usually silences genes. Heterochromatin forms in specific regions like centromeres and telomeres and is important for chromosome function and stability.
This document summarizes information about bacteriophage vectors. It discusses how bacteriophages infect and replicate within bacterial cells, and how this process can be utilized to clone DNA fragments. Specifically, it describes commonly used phage vectors like lambda phage, M13 phage, and P1 phage. Lambda phage vectors can be insertion or replacement vectors, and allow cloning of fragments up to 24kb. M13 phage is a filamentous phage that can deliver single-stranded genomes up to 6.4kb. P1 phage can clone very large fragments up to 1000kb. In conclusion, bacteriophage vectors are useful tools for cloning large DNA fragments.
The document summarizes various genetic techniques including PCR, restriction mapping, the human genome project, in situ hybridization, and cloning the gene responsible for alkaptonuria. It provides an example of how PCR, genomic libraries, DNA sequencing, and other methods were used to clone the HGO gene involved in alkaptonuria. Ethical considerations are discussed around using genetic testing to predict late-onset genetic disorders in fetuses.
Wiki glossary epigenetic control of gene expressionBárbara Pérez
This document is a glossary defining terms related to epigenetics and gene expression. It defines over 80 key terms concisely, including definitions for adenosine triphosphate, alleles, autosomes, Barr body, bisulfite sequencing, blastocyst, boundary element, central dogma, chromatin, chromosome, cytosine methylation, DNA methylation, epigenetic reprogramming, epigenetics, embryonic stem cells, euchromatin, gene, genomic imprinting, histones, homologous chromosomes, imprinted genes, meiosis, mitosis, mRNA, nucleosome, placenta, and pluripotent cells. The glossary provides succinct yet informative definitions for fundamental concepts in molecular biology
This document summarizes recent evidence that the arrangement of chromosomes, gene loci, and nuclear bodies within the cell nucleus is not random but rather exhibits spatial organization that influences gene expression and nuclear processes. Techniques such as fluorescence in situ hybridization and chromosome conformation capture have provided insights into the positioning of chromosomes and genes relative to nuclear landmarks. Chromosomes occupy distinct territories within the nucleus and preferentially localize near the nuclear interior or periphery depending on their gene content. Association with nuclear subcompartments such as the nuclear lamina, nuclear pores, nucleoli, and polycomb bodies can impact the transcriptional state of genes and chromatin domains. Advances in genome-wide and time-lapse imaging approaches are helping to further characterize nuclear organization
This document discusses cloning vectors and plasmid-based cloning vectors. It provides information on plasmids, their structure, replication, and how they are used as cloning vectors. Some key points:
- Plasmids are small DNA molecules that can replicate independently of the bacterial chromosome and are often used as cloning vectors. They exist naturally in bacteria.
- Plasmids can have different structures depending on whether their DNA forms covalently closed circles or open circles. Supercoiled plasmids separate from open circles during electrophoresis.
- Plasmids can replicate in bacteria and confer phenotypes to their host like antibiotic resistance. They are categorized based on properties like being conjugative or copy number.
-
SYNTHETIC CHROMOSOME PLATFORMs IN PLANTS: CONCEPTS & APPLICATIONskundan Jadhao
This document outlines a seminar on synthetic/artificial chromosomes. It begins by discussing the need for synthetic chromosomes due to limitations of traditional genetic engineering approaches. It then describes various methods that have been used to develop artificial chromosomes, including the bottom-up and top-down methods. Several case studies are presented where telomere-mediated truncation was used to produce engineered minichromosomes in plants with endogenous centromeres. The document concludes by discussing ways that engineered minichromosomes can be amended in planta, such as through site-specific recombination systems and zinc finger nucleases.
This document discusses P1 vectors, which are cloning vectors derived from the P1 bacteriophage. P1 can transfer DNA between bacterial cells through transduction. The document describes:
1) The construction of P1 vectors pNS358 and pNS582, which contain P1 packaging and replication elements to clone and propagate large DNA fragments.
2) How the P1 system was used to clone DNA fragments up to 100 kb by packaging ligated vector/insert concatemers into phage heads.
3) Applications of P1 vectors including the creation of P1-derived artificial chromosomes (PACs) which can accommodate relatively large DNA fragments for cloning and mapping genomes.
Dna content,c value paradox, euchromatin heterochromatin, banding patternArchanaSoni3
DNA content refers to the amount of DNA in an organism's haploid chromosomes. It varies greatly between organisms, with eukaryotes generally having more DNA than prokaryotes. The amount of DNA does not always correlate with an organism's complexity, known as the C-value paradox. This is because eukaryotic DNA contains large amounts of non-coding repetitive sequences. Chromatin exists in two forms - euchromatin, which is less condensed and permits gene expression, and heterochromatin, which is highly condensed and usually silences genes. Heterochromatin forms in specific regions like centromeres and telomeres and is important for chromosome function and stability.
Plasmids are commonly used as cloning vectors. They are circular DNA molecules that can replicate independently of the bacterial chromosome. Many plasmid cloning vectors were constructed from bacterial plasmids and contain an origin of replication, selectable marker like antibiotic resistance, and a multiple cloning site to insert DNA fragments. Successful clones can be identified by techniques like blue-white screening where expression of a gene like lacZ allows colonies to be screened blue or white.
This document discusses the history and properties of plasmids. Plasmids are extrachromosomal DNA molecules that are able to replicate independently of a cell's chromosomal DNA. They were first observed in bacteria in the early 1950s and play important roles in processes like antibiotic resistance and virulence. The document outlines the early studies that helped characterize plasmids and describes some of their key properties, such as their circular structure and ability to exist in different conformations. It also discusses how plasmids are used as cloning vectors to amplify genes and produce proteins for applications in research, medicine, and agriculture.
This document summarizes research on the DNA translocase SpoIIIE in Bacillus subtilis. The key findings are:
1) SpoIIIE transports DNA directionally from the mother cell to the forespore during sporulation.
2) The g-domain of SpoIIIE is necessary for establishing directionality of DNA transport in vivo, but not for mechanical translocation in vitro.
3) SpoIIIE recognizes specific DNA sequences called SRS that are highly skewed on the B. subtilis chromosome, similar to how the related protein FtsK recognizes KOPS sequences. Interaction with SRS sequences regulates the direction of SpoIIIE-mediated DNA transport.
MULTIPLE CLONING SITE and BACTERIOPHAGE LAMBDA (λ - phage ) Amany Elsayed
This document summarizes key aspects of cloning vectors and techniques. It discusses multiple cloning sites that contain unique restriction enzyme sites used to insert DNA fragments. Both plasmid vectors and bacteriophage lambda can be used for cloning, with lambda able to accommodate larger inserts up to 25kb. The document compares features of plasmid and lambda cloning, such as circular vs. linear DNA, transformation vs. transfection, and plating methods. It also explains the transformation process, including factors that influence efficiency like calcium chloride treatment and host bacterial strain.
Cloning vectors are DNA molecules that can carry foreign DNA fragments into recipient cells. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and phagemids. Key components of vectors include an origin of replication to replicate in the host cell, selectable markers like antibiotic resistance genes to identify transformed cells, and restriction enzyme sites to insert foreign DNA. Vectors derived from bacteria like E. coli plasmid pBR322 and bacteriophage λ are widely used to clone DNA in prokaryotic cells, while plant Ti plasmids and shuttle vectors allow cloning in both prokaryotic and eukaryotic cells.
The document provides information about bacterial transformation. It describes that transformation is the process by which bacteria take up extracellular DNA from their environment. Frederick Griffith first discovered transformation in 1928 while working with pneumococcus bacteria. His experiments showed that a non-virulent rough form could be transformed into a virulent smooth form by DNA from a heat-killed smooth strain. Later experiments by Avery, Macleod and McCarty demonstrated that DNA is the transforming principle and genetic material of bacteria. The document then discusses various methods of bacterial transformation including chemical and physical methods like electroporation and use of calcium chloride. It also explains the molecular mechanism of transformation involving DNA binding, penetration, synapsis formation and integration into the bacterial chromosome.
Hello everyone, I am Dr. Ujwalkumar Trivedi, Head of Biotechnology Department at Marwadi University Rajkot. I teach Molecular Biology to the students of M.Sc. Microbiology and Biotechnology.
The current presentation talks about replication and partition mechanism of plasmid. The later part of the presentation describes "Theta Model" and "Rolling Circle Model" of replication.
The document summarizes key developments in the structural characterization of G protein-coupled receptors (GPCRs), beginning with the 1975 structure of bacteriorhodopsin determined by Henderson and Unwin, showing its 7 transmembrane alpha helical structure. It then discusses important milestones such as the 1983 cloning of bovine rhodopsin, the first GPCR cDNA; the 1986 cloning of the beta-2 adrenergic receptor, the first non-sensory GPCR; the 1988 cloning of the 5-HT1A receptor, the first "orphan" GPCR to be deorphanized; and the 1991 crystal structure of rhodopsin, the first GPCR structure. The document concludes with the
The document summarizes key developments in the structural characterization of G protein-coupled receptors (GPCRs), beginning with the 1975 structure of bacteriorhodopsin determined by Henderson and Unwin, showing its 7 transmembrane alpha helical structure. It then discusses important milestones such as the 1983 cloning of bovine rhodopsin, the first GPCR cDNA sequence, the 1986 cloning of the beta-2 adrenergic receptor, the first non-sensory GPCR, and the 1991 crystal structure of rhodopsin, the first GPCR structure determined. The document concludes with the 2007 crystal structure of the beta-2 adrenergic receptor bound to G protein and ligand, and the 2012 Nobel Prize
The document summarizes phagemid and bacterial artificial chromosome (BAC) vectors. It describes that phagemid vectors are plasmids that contain both plasmid and phage origins of replication. Specifically, it discusses the features of pBluescript II phagemid vectors, including their polylinker and RNA polymerase promoter sequences. It also describes how pBluescript II phagemid vectors can produce blue or white colonies depending on insert presence. The document then explains that BAC vectors are low-copy plasmids that can hold up to 300kb DNA fragments. Examples of BAC vectors like pBAC108L and pBeloBAC11 are provided, with details about their replication origin and partitioning functions.
Vectors part 1 | molecular biology | biotechnologyatul azad
This document discusses various types of plasmid and bacteriophage vectors used for cloning DNA fragments. It describes the features and selection methods of commonly used vectors like pBR322, pUC18, pGEM3Z, lambda phage and M13 phage vectors. Plasmid vectors like pBR322 are advantageous for having small size, high copy number and antibiotic resistance markers. Lambda phage vectors can accommodate larger inserts compared to plasmids and allow easy screening of recombinant phages. M13 vectors are useful for obtaining single-stranded DNA copies for sequencing.
Relaxed plasmids & Regulation of copy numbersalvia16
Plasmids are classified as either stringent or relaxed based on their copy number in bacterial cells. Stringent plasmids exist in low copy numbers (<100 copies/cell) and rely on the bacterial genome for replication and segregation. Relaxed plasmids exist in high copy numbers (>100 copies/cell) and replicate independently of the bacterial genome. Relaxed plasmids include ColE1, which uses an antisense RNA mechanism to regulate its copy number based on plasmid concentration in the cell.
Bacteriophage lambda is a double-stranded DNA virus that can be engineered as an insertion vector for cloning foreign DNA fragments. Lambda has a 48.5 kb genome that is linear with single-stranded overhangs. Insertion vectors like GT10 and GT11 contain a unique restriction site, like EcoRI, within a gene where foreign DNA can be inserted. This inactivates genes required for lysogeny, allowing selection of recombinant phage by plaquing on lysogenic E. coli strains. Larger DNA fragments can be cloned using substitution vectors.
Lectut btn-202-ppt-l3. gene cloning and plasmid vectors (1)Rishabh Jain
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of other vector types were also constructed for different applications, such as bacteriophages, cosmids, YACs, BACs, and artificial chromosomes.
Vectors such as plasmids, lambda phage, cosmids, and yeast artificial chromosomes can be used to carry DNA fragments into host cells. Cosmids are hybrid vectors derived from plasmids that contain lambda phage cos sites, allowing them to carry 37-52 kilobase pairs of DNA. Yeast artificial chromosomes are capable of carrying large DNA fragments of up to 2 megabases but have low transformation efficiency. Plasmids are extrachromosomal DNA molecules that are important tools for cloning genes and transferring traits such as antibiotic resistance between bacteria.
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of vectors were also developed using bacteriophages, cosmids, artificial chromosomes, and more to clone larger fragments.
Plasmid is a double stranded, circular extra chromosomal DNA of bacterium. It is used in recombinant DNA experiments to clone genes from other organisms and make large quantities of their DNA. Plasmid can be transferred between same species or between different species. Size of plasmids range from 1-1000 kilo base pairs. Plasmids are part of mobilomes (total of all mobile genetic elements in a genome) like transposons or prophages and are associated with conjugation. Even the largest plasmids are considerably smaller than the chromosomal DNA of the bacterium, which can contain several million base pairs.
Este documento proporciona una guía sobre el uso de Microsoft Outlook. Explica cómo configurar cuentas de correo electrónico, administrar correos, contactos, tareas, citas y reuniones. También describe las características y funciones generales de Outlook como enviar y recibir correo, sincronizar con otros dispositivos, y realizar búsquedas.
Plasmids are commonly used as cloning vectors. They are circular DNA molecules that can replicate independently of the bacterial chromosome. Many plasmid cloning vectors were constructed from bacterial plasmids and contain an origin of replication, selectable marker like antibiotic resistance, and a multiple cloning site to insert DNA fragments. Successful clones can be identified by techniques like blue-white screening where expression of a gene like lacZ allows colonies to be screened blue or white.
This document discusses the history and properties of plasmids. Plasmids are extrachromosomal DNA molecules that are able to replicate independently of a cell's chromosomal DNA. They were first observed in bacteria in the early 1950s and play important roles in processes like antibiotic resistance and virulence. The document outlines the early studies that helped characterize plasmids and describes some of their key properties, such as their circular structure and ability to exist in different conformations. It also discusses how plasmids are used as cloning vectors to amplify genes and produce proteins for applications in research, medicine, and agriculture.
This document summarizes research on the DNA translocase SpoIIIE in Bacillus subtilis. The key findings are:
1) SpoIIIE transports DNA directionally from the mother cell to the forespore during sporulation.
2) The g-domain of SpoIIIE is necessary for establishing directionality of DNA transport in vivo, but not for mechanical translocation in vitro.
3) SpoIIIE recognizes specific DNA sequences called SRS that are highly skewed on the B. subtilis chromosome, similar to how the related protein FtsK recognizes KOPS sequences. Interaction with SRS sequences regulates the direction of SpoIIIE-mediated DNA transport.
MULTIPLE CLONING SITE and BACTERIOPHAGE LAMBDA (λ - phage ) Amany Elsayed
This document summarizes key aspects of cloning vectors and techniques. It discusses multiple cloning sites that contain unique restriction enzyme sites used to insert DNA fragments. Both plasmid vectors and bacteriophage lambda can be used for cloning, with lambda able to accommodate larger inserts up to 25kb. The document compares features of plasmid and lambda cloning, such as circular vs. linear DNA, transformation vs. transfection, and plating methods. It also explains the transformation process, including factors that influence efficiency like calcium chloride treatment and host bacterial strain.
Cloning vectors are DNA molecules that can carry foreign DNA fragments into recipient cells. Common types of cloning vectors include plasmids, bacteriophages, cosmids, and phagemids. Key components of vectors include an origin of replication to replicate in the host cell, selectable markers like antibiotic resistance genes to identify transformed cells, and restriction enzyme sites to insert foreign DNA. Vectors derived from bacteria like E. coli plasmid pBR322 and bacteriophage λ are widely used to clone DNA in prokaryotic cells, while plant Ti plasmids and shuttle vectors allow cloning in both prokaryotic and eukaryotic cells.
The document provides information about bacterial transformation. It describes that transformation is the process by which bacteria take up extracellular DNA from their environment. Frederick Griffith first discovered transformation in 1928 while working with pneumococcus bacteria. His experiments showed that a non-virulent rough form could be transformed into a virulent smooth form by DNA from a heat-killed smooth strain. Later experiments by Avery, Macleod and McCarty demonstrated that DNA is the transforming principle and genetic material of bacteria. The document then discusses various methods of bacterial transformation including chemical and physical methods like electroporation and use of calcium chloride. It also explains the molecular mechanism of transformation involving DNA binding, penetration, synapsis formation and integration into the bacterial chromosome.
Hello everyone, I am Dr. Ujwalkumar Trivedi, Head of Biotechnology Department at Marwadi University Rajkot. I teach Molecular Biology to the students of M.Sc. Microbiology and Biotechnology.
The current presentation talks about replication and partition mechanism of plasmid. The later part of the presentation describes "Theta Model" and "Rolling Circle Model" of replication.
The document summarizes key developments in the structural characterization of G protein-coupled receptors (GPCRs), beginning with the 1975 structure of bacteriorhodopsin determined by Henderson and Unwin, showing its 7 transmembrane alpha helical structure. It then discusses important milestones such as the 1983 cloning of bovine rhodopsin, the first GPCR cDNA; the 1986 cloning of the beta-2 adrenergic receptor, the first non-sensory GPCR; the 1988 cloning of the 5-HT1A receptor, the first "orphan" GPCR to be deorphanized; and the 1991 crystal structure of rhodopsin, the first GPCR structure. The document concludes with the
The document summarizes key developments in the structural characterization of G protein-coupled receptors (GPCRs), beginning with the 1975 structure of bacteriorhodopsin determined by Henderson and Unwin, showing its 7 transmembrane alpha helical structure. It then discusses important milestones such as the 1983 cloning of bovine rhodopsin, the first GPCR cDNA sequence, the 1986 cloning of the beta-2 adrenergic receptor, the first non-sensory GPCR, and the 1991 crystal structure of rhodopsin, the first GPCR structure determined. The document concludes with the 2007 crystal structure of the beta-2 adrenergic receptor bound to G protein and ligand, and the 2012 Nobel Prize
The document summarizes phagemid and bacterial artificial chromosome (BAC) vectors. It describes that phagemid vectors are plasmids that contain both plasmid and phage origins of replication. Specifically, it discusses the features of pBluescript II phagemid vectors, including their polylinker and RNA polymerase promoter sequences. It also describes how pBluescript II phagemid vectors can produce blue or white colonies depending on insert presence. The document then explains that BAC vectors are low-copy plasmids that can hold up to 300kb DNA fragments. Examples of BAC vectors like pBAC108L and pBeloBAC11 are provided, with details about their replication origin and partitioning functions.
Vectors part 1 | molecular biology | biotechnologyatul azad
This document discusses various types of plasmid and bacteriophage vectors used for cloning DNA fragments. It describes the features and selection methods of commonly used vectors like pBR322, pUC18, pGEM3Z, lambda phage and M13 phage vectors. Plasmid vectors like pBR322 are advantageous for having small size, high copy number and antibiotic resistance markers. Lambda phage vectors can accommodate larger inserts compared to plasmids and allow easy screening of recombinant phages. M13 vectors are useful for obtaining single-stranded DNA copies for sequencing.
Relaxed plasmids & Regulation of copy numbersalvia16
Plasmids are classified as either stringent or relaxed based on their copy number in bacterial cells. Stringent plasmids exist in low copy numbers (<100 copies/cell) and rely on the bacterial genome for replication and segregation. Relaxed plasmids exist in high copy numbers (>100 copies/cell) and replicate independently of the bacterial genome. Relaxed plasmids include ColE1, which uses an antisense RNA mechanism to regulate its copy number based on plasmid concentration in the cell.
Bacteriophage lambda is a double-stranded DNA virus that can be engineered as an insertion vector for cloning foreign DNA fragments. Lambda has a 48.5 kb genome that is linear with single-stranded overhangs. Insertion vectors like GT10 and GT11 contain a unique restriction site, like EcoRI, within a gene where foreign DNA can be inserted. This inactivates genes required for lysogeny, allowing selection of recombinant phage by plaquing on lysogenic E. coli strains. Larger DNA fragments can be cloned using substitution vectors.
Lectut btn-202-ppt-l3. gene cloning and plasmid vectors (1)Rishabh Jain
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of other vector types were also constructed for different applications, such as bacteriophages, cosmids, YACs, BACs, and artificial chromosomes.
Vectors such as plasmids, lambda phage, cosmids, and yeast artificial chromosomes can be used to carry DNA fragments into host cells. Cosmids are hybrid vectors derived from plasmids that contain lambda phage cos sites, allowing them to carry 37-52 kilobase pairs of DNA. Yeast artificial chromosomes are capable of carrying large DNA fragments of up to 2 megabases but have low transformation efficiency. Plasmids are extrachromosomal DNA molecules that are important tools for cloning genes and transferring traits such as antibiotic resistance between bacteria.
The document discusses various types of plasmid vectors and cloning vectors used for gene cloning. It describes the key properties required for a vector, including autonomous replication, small size, selectable markers, and restriction enzyme sites. Some examples of early plasmid vectors discussed are pSC101, ColE1, and pBR322. Later vectors with improved properties include the pUC series, pGEM series, and pET series. A variety of vectors were also developed using bacteriophages, cosmids, artificial chromosomes, and more to clone larger fragments.
Plasmid is a double stranded, circular extra chromosomal DNA of bacterium. It is used in recombinant DNA experiments to clone genes from other organisms and make large quantities of their DNA. Plasmid can be transferred between same species or between different species. Size of plasmids range from 1-1000 kilo base pairs. Plasmids are part of mobilomes (total of all mobile genetic elements in a genome) like transposons or prophages and are associated with conjugation. Even the largest plasmids are considerably smaller than the chromosomal DNA of the bacterium, which can contain several million base pairs.
Este documento proporciona una guía sobre el uso de Microsoft Outlook. Explica cómo configurar cuentas de correo electrónico, administrar correos, contactos, tareas, citas y reuniones. También describe las características y funciones generales de Outlook como enviar y recibir correo, sincronizar con otros dispositivos, y realizar búsquedas.
Le pouvoir des signes
chapitre 9 -des artistes militants
Page 189
Images du sourd dans l'audiovisuel.
Guy JOUANNET - Rédacteur-documentaliste cinéma, Charenton (94)
Le pouvoir des signes
Page 208
"La langue des signes française de 1978 à nos jours."
Guy Bouchauveau - Attaché scientifique sourd, Cité des sciences et de l'industrie, Paris.
Page 215
"Les sourds aux Etats-Unis après Laurent Clerc."
Harlan LANE ^ph D., professeur de psychologie, North Eastern University, Boston USA.
Page 222
"Construire l'espace des sourds"
Myriam SEILLER - Urbaniste, interprète en LSF.
Pierre FABRE - Sociologue, économiste.
L’Autorité des marchés financiers (AMF) publie un état des lieux des pratiques en matière de stratégie et communication actionnariale des sociétés cotées. Première étude de cet ordre menée par les services de l’AMF, elle dresse le bilan et les bonnes pratiques étudiées auprès de 80 émetteurs. Fort de ces enseignements, le régulateur complète par ailleurs sa doctrine avec la publication d’une position et d’une recommandation.
La Villa Savoye de Le Corbusier aplicó los cinco principios de la arquitectura racionalista: se elevó sobre pilotis, tuvo planta libre, fachada libre, ventanas longitudinales y azotea jardín. El edificio se componía de tres niveles conectados por una escalera y rampa. Cada nivel tenía funciones diferentes como servicios, áreas privadas y solárium. Los materiales y estructura independiente de la forma maximizaron la funcionalidad del espacio.
México tiene una gran diversidad de ecosistemas naturales, incluyendo desiertos, bosques de coníferas y encino, selvas tropicales húmedas y secas. Los bosques de pino y encino albergan una gran diversidad de especies, pero se encuentran amenazados por la deforestación. México también posee una gran variedad de ecosistemas acuáticos como praderas de pastos marinos.
This document provides instructions for creating, managing, and editing Discreet Work Requirements (DWRs) and associated work instructions in a software system. It outlines steps for creating events and statements of work to which DWRs can be added. It also describes how to generate work instructions from DWRs and manage parts lists. The guide is intended to help users efficiently navigate relevant software menus and pages to complete tasks.
Discurso traspaso de poderes municipalidad corredoresCarlos Viales
Este discurso de toma de posesión del nuevo alcalde expresa su agradecimiento por ser elegido para el cargo. Reconoce los grandes retos que enfrenta y la necesidad de trabajar en cooperación con el Concejo Municipal y otras autoridades. Anuncia que iniciará un estudio para actualizar el Manual de Puestos de la municipalidad y mejorar las condiciones laborales de los funcionarios, con el fin de prestar un mejor servicio a los ciudadanos.
The document discusses Project Management Institute Pearl City Chapter's Student Leadership Competency Building initiative. The initiative aims to develop leadership skills in students aged 12-22 so they can become industry and nation ready leaders. It involves establishing Student Leadership Advisory Councils at partner academic institutions to implement training programs focused on leadership competencies. The goal is to help students gain the skills needed to contribute to the growth of the nation and industry.
Alegría de Evangelizar y la renovación de la escuela patricia0306
El documento propone una renovación de la escuela católica para enfrentar los desafíos del tiempo actual. Sugiere centrarse en lo esencial, crear un clima de alegría y fraternidad con Jesús, y escuchar a los estudiantes. También recomienda no renunciar a la identidad cristiana y ofrecer la propuesta educativa en un lenguaje accesible para todos. El objetivo es enseñar a pensar críticamente y derramar la vida de Jesús en las nuevas generaciones.
Jerry is a restaurant manager known for his positive attitude. When asked how he maintains this outlook, Jerry explains that every day he chooses whether to have a good or bad mood. He also chooses to learn from bad experiences rather than see himself as a victim. Jerry was later the victim of an armed robbery where he was shot. Despite his injuries, Jerry chose to live and maintained his positive attitude during his long recovery process. He credits his choice to stay positive with helping him survive.
This study investigates what happens to proteins associated with DNA during chromosome translocation in Bacillus subtilis sporulation. Using fluorescent protein fusions and a mutant that forms two forespores, the authors show that RNA polymerase, chromosome remodeling proteins, and transcription factors are stripped off the chromosome as it is translocated into the forespores. Specifically, they demonstrate that a TetR-GFP fusion bound to an operator array is efficiently removed from the translocating DNA. Additionally, in vitro experiments indicate that the ATPase domain of SpoIIIE can displace RNA polymerase from DNA. These results suggest that SpoIIIE translocates naked DNA and strips associated proteins during chromosome transport, which may play a role in reprogramming gene
1) Bacterial cell division involves coordinating the segregation of replicated DNA and division of cytoplasmic material to generate progeny with identical genetic material. This requires spatial and temporal coordination of events.
2) The divisome, consisting of 10-15 proteins including FtsZ, assembles at midcell after DNA replication begins to assist in DNA separation and synthesize a new cell wall to divide the daughter cells.
3) There are multiple mechanisms that regulate divisome assembly to prevent it from occurring too early or in the same location as segregating chromosomes.
This document summarizes recent research on the role of chromosome architecture in bacterial cellular organization. It discusses how chromosome structure encodes spatial information that directs processes like chromosome compaction, replication initiation, and segregation. Specifically, it describes how in Caulobacter, the positioning of the origin region near the cell pole facilitates cell cycle regulation. It also explains how the nucleoid provides a scaffold for partitioning systems to segregate chromosomal loci and other cargos through the cell. Finally, it discusses mechanisms in several bacteria by which dynamic changes to chromosome architecture couple replication and segregation to proper placement of the cell division machinery.
FtsK is a DNA translocase in E. coli that is involved in chromosome dimer resolution and segregation. Through single-molecule experiments tracking FtsK movement on DNA substrates containing the E. coli chromosome region near the dif site, the authors identified four zones where FtsK frequently reverses direction of translocation. Analysis of these zones identified the DNA sequence motif GNGNAGGG as a candidate sequence that causes FtsK to pause and reverse direction upon encountering it from the 3' end of the G-rich strand. FtsK paused for an average of 1.0 seconds at these reversal sites.
This document provides evidence that common machinery is utilized by the early and late RNA localization pathways in Xenopus oocytes. It presents four key findings: 1) Early and late pathway RNAs require the same short sequence motifs for localization. 2) Competition assays show early and late RNAs compete for common localization factors in vivo. 3) A late localization factor, Vg RBP/Vera, binds specifically to localization elements of early pathway RNAs. 4) Confocal imaging reveals early RNAs associate with microtubules, suggesting transport plays a role in both pathways. Together, these findings suggest the early and late pathways share basal localization factors throughout oogenesis.
RNA localization to the Balbiani body in Xenopus oocytes is regulated by the energy state of the cell and is facilitated by kinesin II. The rate of RNA accumulation in the Balbiani body depends on temperature and intracellular ATP concentration - increasing ATP concentration doubles the localization rate. Inhibition of kinesin II reduces RNA localization to the Balbiani body, and the Xcat-2 RNA recruits kinesin II, indicating it plays a role in this process. The energy state of the cell regulates the rate of RNA transport to the Balbiani body, which involves kinesin II to some extent.
This document summarizes research characterizing the mumps virus nucleocapsid-binding domain (NBD) protein via fluorescence spectroscopy and circular dichroism. Researchers created a variant of the mumps NBD protein called F366W by introducing a tryptophan mutation using site-directed mutagenesis. This allowed stability measurements using fluorescence spectroscopy. The mutated protein was expressed in E. coli and purified. Fluorescence spectroscopy and circular dichroism were used to monitor the protein's tertiary structure as it was subjected to thermal and chemical denaturation conditions. Results from these experiments provided insights into the stability and folding of the mumps NBD protein.
Replication in prokaryotes involves three main stages - initiation, elongation, and termination. Initiation begins at a specific origin site and involves unwinding and formation of a prepriming complex. Elongation then begins, with leading strand synthesis occurring continuously and lagging strand synthesis occurring via Okazaki fragments. Replication is completed when the replication forks from opposing directions meet up at termination sites. The entire replication process in E. coli takes about 40 minutes.
The distribution of callose synthase, cellulose synthase, and sucrose synthase in tobacco pollen tubes is controlled by actin filaments and microtubules in different ways. Cellulose synthase is present along the entire length of pollen tubes, with higher concentration at the apex, while callose synthase is located in the apex and around callose plugs. Actin filaments and endomembrane dynamics control the distribution of both enzymes by transporting them via Golgi bodies and vesicles. Microtubules control the positioning of callose synthase in distal regions and around plugs, but only partially influence cellulose synthase distribution. Sucrose synthase binds to actin filaments and provides UDP-
This doctoral research used single-molecule imaging techniques to study DNA replication in live E. coli cells. Endogenous replication proteins like β2-clamps were labeled with fluorescent proteins using genome engineering. Observations found β2-clamps accumulate on DNA after initiation and remain bound for minutes. Experiments also examined how replisomes encounter natural Tus-Ter roadblocks and determined replication speed decreases but is not stalled. Further work aimed to study primase dynamics and localization of Tus proteins during the cell cycle but faced challenges in strain engineering. The research demonstrated applications of in vivo single-molecule imaging while also highlighting ongoing difficulties in the field.
This document summarizes our current understanding of DNA replication in archaea. It discusses how archaeal replication is more similar to eukaryotic replication than bacterial replication, though a simpler version. A key point is that while putative origins of replication have been identified in archaea based on genomic analysis, an unambiguous chromosomal origin has not been found. The document also provides overviews of DNA replication processes in general and the initiation phase more specifically to help frame the discussion of archaeal replication.
This document summarizes a study examining the formation and function of the contractile ring involved in polar body extrusion in the surf clam Spisula. The study found that:
1) The metaphase peripheral aster spreads along the egg cortex in an umbrella-like pattern, leaving a microtubule-poor center.
2) During anaphase, the aster disassembles as a cortical F-actin ring forms that matches the location, size, and pattern of the previous aster.
3) Inhibiting F-actin or myosin blocked polar body formation, while disrupting or stabilizing asters prevented proper ring and polar body formation, supporting the hypothesis that aster spreading
The document describes the process of purifying the elongation factor LepA/EF4 protein from E. coli. The gene for EF4 was transformed into E. coli cells using a plasmid. The cells were then lysed using sonication and the EF4 protein was purified from the cell lysate using affinity chromatography and its hexahistidine tag. The concentration of the purified EF4 protein was determined to be 0.57 μg/μL using a Bradford assay and its molecular weight was found to be ~69 kDa by SDS-PAGE. Secondary and tertiary structural analysis using circular dichroism and fluorescence spectroscopy yielded thermodynamic values for EF4 protein denaturation.
1) Researchers have discovered that the giant Mimivirus, which was originally mistaken for a bacterium, can be infected by a smaller virus called Sputnik.
2) When amoeba cells were inoculated with both Mimivirus and Sputnik, both viruses were able to multiply inside the "virus factory" that Mimivirus builds in the amoeba cells during its lytic infection cycle.
3) Sputnik reproduces faster than Mimivirus inside the virus factory, with new Sputnik virions emerging after 6 hours while new Mimivirus virions only appear after 8 hours. Infection with both viruses decreases the yield of infective Mimivirus vir
This study examines how applied hydrodynamic drag force affects the contractions of live Vorticella by impeding their contractions in a microfluidic channel. As the stall force increased by changing the flow rate and viscosity of the solution, the contracted stalk length increased and maximum contraction speed decreased, while contractions took longer. The time lag in contraction between the zooid and stalk also increased. This implies that the stalk cannot contract until it develops enough force to overcome the stall force. As stall force increased, relaxations took longer and the stalk resumed contraction after the force was removed, showing that the spasmoneme retains contractile force but the stall force extends the stalk.
Exchange of genes between two DNA molecules to form new combinations of genes on a chromosome
contributes to a population’s genetic diversity (source of variation in evolution)
Recombination is more likely than mutation to be beneficial
Less likely destroy a gene's function
May bring together combinations of genes
This document summarizes the key steps in the cycle of actin remodeling at the cell surface. It discusses (1) initiation of actin polymerization through uncapping of barbed ends or nucleation by Arp2/3 or formins, (2) elongation through barbed end polymerization and regulation by capping proteins, (3) termination through capping or stabilization, (4) branching through Arp2/3, (5) crosslinking into networks, (6) contraction through myosins and cargo transport, (7) membrane attachment, and (8) disassembly through severing by gelsolin or ADF/cofilin and capping of barbed ends. The cycle is regulated
Restriction enzymes were discovered in the 1960s and cut DNA at specific recognition sequences. There are five types of restriction enzymes that differ in subunit composition and sequence recognition. Restriction digestion leaves sticky or blunt ends that can be joined by DNA ligase. Transformation involves the uptake and recombination of extracellular DNA into bacterial cells through natural competence, chemical treatments, or electroporation. Genomic libraries contain fragmented, cloned DNA that represent an organism's entire genome.
The document summarizes experiments studying the movement and directionality of the FtsK translocase protein on DNA molecules. Key findings include:
1) FtsK moves rapidly at 5 kilobases per second and can work against forces up to 60 piconewtons.
2) FtsK forms loops of DNA as it translocates and can reverse direction without dissociating from DNA.
3) When bound to lambda DNA, FtsK consistently moved in the same direction, toward the terminus region as predicted, even when the DNA was inverted.
This summary describes a study that uses fluorescence microscopy to visualize the actin cytoskeleton during sperm individualization in Drosophila melanogaster. The researchers develop a simple assay using rhodamine-phalloidin to label F-actin in the individualization complex (IC), which packages each sperm cell during development. Using this assay, they analyze male-sterile mutants and identify four classes based on defects in IC assembly and movement. Class 1 mutants block IC assembly, class 2 mutants allow assembly but not movement, class 3 mutants allow initial movement but the IC then breaks down, and class 4 mutants allow movement but with an altered morphology. This assay provides insights into the cellular mechanisms of sperm individualization and how different mutations can disrupt this
Similar to Kennedy_et_al-2008-Molecular_Microbiology (20)