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This document summarizes a study on cloning and analyzing glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes from different plant species. The researcher amplified GAPDH genes from Oxalis corniculata, Plectranthus amboinicus, and Myrtaceae psidium via PCR. They then cloned the genes into E. coli and sequenced them using Sanger sequencing. The goal was to compare the GAPDH sequences to analyze conserved amino acids in the active site and differences elsewhere, helping understand GAPDH protein function and evolution across plants. Preliminary results were obtained but not detailed.
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Genome editing
This document discusses genome editing techniques. It begins by defining genomes and how they consist of DNA or RNA that contains both coding and non-coding regions. It then discusses several methods of genome editing including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas system. Each method uses engineered nucleases to introduce targeted double-strand breaks in DNA, allowing the cell's repair mechanisms to modify the genome. The CRISPR-Cas system was selected as the breakthrough of the year in 2015 due to its simplicity, efficiency and precision for genome editing applications.
TALENs: A WIDELY APPLICABLE TECHNOLOGY FOR TARGETED GENOME EDITING
A seminar presentation on TALENs a tool most widely used in genome editing for building up desired traits in crop plants
Proposal final
This study aims to clone and analyze GAPDH genes from various plant species to compare amino acid sequences related to catalytic function. Previous work successfully cloned GAPDH from Oxalis corniculata and Plectranthus amboinicus but not Myrtaceae psidium. The current study will continue work on the cloned samples through midipreps, restriction enzyme digestion and sequencing. Sequence data will undergo bioinformatics analysis to compare conserved amino acids of the GAPDH protein between plant species. Results could provide new genetic information published in GenBank and further the understanding of energy production pathways in plants.
Final ppt
This presentation provides an overview of genome editing. It defines genome editing as a technique used to modify DNA within a cell precisely and efficiently using engineered nucleases. The presentation discusses different tools for genome editing including meganucleases, zinc finger nucleases, TALENs, and CRISPR. It explains how these tools work by creating cuts in DNA at specific sequences, and how cells naturally repair this using homologous directed repair or non-homologous end joining. Applications of genome editing discussed include gene therapy, modifying crops and animals, disease treatment, and ecological control.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)
clustered regularly interspaced short palindromic repeats is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria. Now CRISPR use as genome editing tool in different Plant Breeder to manipulate the DNA of the crop
Genome editing techniques
Genome editing techniques allow DNA to be inserted, deleted, modified or replaced in the genome of a living organism. Four families of engineered nucleases have been used for genome editing: meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR/Cas9 system. Each system uses a different mechanism for recognizing and cutting DNA at specific locations to modify genes. The CRISPR/Cas9 system has become widely used due to its ease of design and lower off-target effects compared to other techniques.
Progress and prospects in plant genome editing
The document summarizes a seminar on plant genome editing tools. It discusses various tools for targeted genome manipulation including zinc finger nucleases, TALENs, and CRISPR/Cas9. It provides examples of each tool being used to generate disease resistance in crops like rice and wheat. It also discusses factors like design, efficiency, and off-target effects of the different tools. Case studies demonstrate using these tools to edit susceptibility genes in rice for bacterial blight resistance and three MLO genes in wheat for powdery mildew resistance.
Gene Editing: An Essential Tool For Plant Breeding
Gene editing is a type of genetic engineering that modifies an organism's DNA by deleting, inserting or replacing parts of the genome. The document discusses several molecular "scissors" or nucleases that can make targeted cuts in DNA, including meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas9 system. These nucleases are used to induce double-strand breaks that are then repaired through natural cellular mechanisms, allowing for targeted changes to the genome. The techniques have been applied successfully in several crop plants to develop traits like herbicide resistance and disease resistance.
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Genome editing
This document discusses genome editing techniques. It begins by defining genomes and how they consist of DNA or RNA that contains both coding and non-coding regions. It then discusses several methods of genome editing including zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas system. Each method uses engineered nucleases to introduce targeted double-strand breaks in DNA, allowing the cell's repair mechanisms to modify the genome. The CRISPR-Cas system was selected as the breakthrough of the year in 2015 due to its simplicity, efficiency and precision for genome editing applications.
TALENs: A WIDELY APPLICABLE TECHNOLOGY FOR TARGETED GENOME EDITING
A seminar presentation on TALENs a tool most widely used in genome editing for building up desired traits in crop plants
Proposal final
This study aims to clone and analyze GAPDH genes from various plant species to compare amino acid sequences related to catalytic function. Previous work successfully cloned GAPDH from Oxalis corniculata and Plectranthus amboinicus but not Myrtaceae psidium. The current study will continue work on the cloned samples through midipreps, restriction enzyme digestion and sequencing. Sequence data will undergo bioinformatics analysis to compare conserved amino acids of the GAPDH protein between plant species. Results could provide new genetic information published in GenBank and further the understanding of energy production pathways in plants.
Final ppt
This presentation provides an overview of genome editing. It defines genome editing as a technique used to modify DNA within a cell precisely and efficiently using engineered nucleases. The presentation discusses different tools for genome editing including meganucleases, zinc finger nucleases, TALENs, and CRISPR. It explains how these tools work by creating cuts in DNA at specific sequences, and how cells naturally repair this using homologous directed repair or non-homologous end joining. Applications of genome editing discussed include gene therapy, modifying crops and animals, disease treatment, and ecological control.
CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)
clustered regularly interspaced short palindromic repeats is a family of DNA sequences found in the genomes of prokaryotic organisms such as bacteria. Now CRISPR use as genome editing tool in different Plant Breeder to manipulate the DNA of the crop
Genome editing techniques
Genome editing techniques allow DNA to be inserted, deleted, modified or replaced in the genome of a living organism. Four families of engineered nucleases have been used for genome editing: meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR/Cas9 system. Each system uses a different mechanism for recognizing and cutting DNA at specific locations to modify genes. The CRISPR/Cas9 system has become widely used due to its ease of design and lower off-target effects compared to other techniques.
Progress and prospects in plant genome editing
The document summarizes a seminar on plant genome editing tools. It discusses various tools for targeted genome manipulation including zinc finger nucleases, TALENs, and CRISPR/Cas9. It provides examples of each tool being used to generate disease resistance in crops like rice and wheat. It also discusses factors like design, efficiency, and off-target effects of the different tools. Case studies demonstrate using these tools to edit susceptibility genes in rice for bacterial blight resistance and three MLO genes in wheat for powdery mildew resistance.
Gene Editing: An Essential Tool For Plant Breeding
Gene editing is a type of genetic engineering that modifies an organism's DNA by deleting, inserting or replacing parts of the genome. The document discusses several molecular "scissors" or nucleases that can make targeted cuts in DNA, including meganucleases, zinc finger nucleases (ZFNs), transcription activator-like effector nucleases (TALENs), and the CRISPR-Cas9 system. These nucleases are used to induce double-strand breaks that are then repaired through natural cellular mechanisms, allowing for targeted changes to the genome. The techniques have been applied successfully in several crop plants to develop traits like herbicide resistance and disease resistance.
Genome Editing Introduction
Genome editing is a method of making specific changes to the DNA of a cell or organism. An enzyme scissors the DNA at a specific sequence, and when this is repaired by the cell, a change or ‘edit’ is made to the sequence.
https://www.creative-biolabs.com/gene-therapy/approaches-to-genome-editing.htm
Genome editing with engineered nucleases
Genome editing uses engineered nucleases to insert, replace or remove DNA from the genome. These nucleases create targeted double-strand breaks which are repaired through natural DNA repair processes, allowing for changes to the genome sequence. Three main engineered nuclease systems for genome editing are ZFNs, TALENs, and CRISPR-Cas9. CRISPR uses a guide RNA and Cas9 nuclease to make precise cuts at targeted DNA sequences for editing. It has advantages over ZFNs and TALENs in being cheaper, easier to design, and more efficient. Genome editing holds promise for applications in crops, medicine, and research.
Methods of screening
This document discusses various methods for screening recombinant clones, including:
1. Blue-white screening using X-gal to detect the presence or absence of insertions disrupting beta-galactosidase activity.
2. Insertional inactivation screening by inserting DNA fragments to disrupt antibiotic resistance or repressor genes.
3. Antibiotic sensitivity screening to detect circularized recombinant plasmids capable of conferring antibiotic resistance.
4. Auxotrophic yeast strain screening using yeast vectors containing selectable marker genes to complement host mutations.
5. Reporter gene assays using enzymes like luciferase, GFP, etc. fused to promoters to identify positive recombinant clones.
Genome editing
This presentation highlights the basics and application of genome editing strategies in plants, strategies to reduce off-target mutation, identification of mutant analysis etc.
P uc vectors
pUC vectors are plasmids derived from pBR322 that have a higher copy number of 500-600 copies per cell. They contain an ampicillin resistance gene for selection, as well as the lacZ' gene containing multiple cloning sites. When a gene of interest is inserted, it disrupts the lacZ' gene, allowing for blue-white screening on media containing IPTG and X-gal to identify recombinant colonies that appear white instead of blue. pUC vectors offer advantages over pBR322 such as high copy number and easy selection, though they cannot accommodate inserts larger than 15kb.
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This document outlines a lecture on microbial genetics. It discusses the genetic engineering processes used to produce human insulin and growth hormone (HGH) in bacteria. For insulin, the human gene is isolated and inserted into bacterial plasmid DNA using restriction enzymes. Transformed bacteria then produce insulin. A similar process is used for HGH, isolating its mRNA from human pituitary gland cells, generating cDNA, and inserting it into bacteria to produce HGH. Lastly, it notes other human hormones can be genetically engineered using these techniques.
Derivatives of pBR322
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.
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Genome editing
Satrupa Das discusses using programmable nucleases like TALENs and CRISPR-Cas9 for genome editing to treat diseases. She describes a study where exon 44 was knocked back into the dystrophin gene of an iPSC line from a Duchenne muscular dystrophy patient using TALENs or Cas9. Over 90% of clones showed targeted integration of the donor template, and sequencing confirmed restoration of the reading frame without other mutations. Differentiated muscle cells from corrected clones expressed dystrophin, demonstrating therapeutic potential of genome editing for Duchenne muscular dystrophy.
Genome Editing Techniques by Kainat Ramzan
Genome technology has revolutionized biological science through techniques of Gene Editing in order to edit any organism's genome.MegNs and zinc-finger nucleases are commonly understood to be used, as is the effector's transcriptional activator-like nucleases. In CRISPR/Cas9, genetic alterations, and gene functionality have become a well-known tool for understanding gene targeting.
ZINC FINGER NUCLEASE TECHNOLOGY
Zinc finger nucleases (ZFNs) allow for highly targeted editing of the genome. ZFNs consist of a DNA-binding domain made of zinc finger proteins and a DNA-cleaving domain. The ZFN pair binds to a target site and creates a double-strand break, which the cell repairs through non-homologous end joining or homologous recombination, enabling gene knockouts or targeted changes. ZFNs work in many cell types and animal models, providing a more efficient alternative to traditional transgenic techniques. They have applications in functional genomics, cell line engineering, and animal model generation.
Crop genome editing using CRISPR
Genome editing uses tools like CRISPR/Cas9 to make precise, targeted changes to DNA in crop plants. The document discusses using CRISPR to edit the SlMlo1 gene in tomato, conferring resistance to powdery mildew. Researchers generated CRISPR/Cas9 plasmids targeting SlMlo1, transformed tomato plants, and identified lines with 48 bp deletions in the gene. Resistant T1 plants lacking the transgene were obtained, named the Tomelo variety. Genome editing provides a faster way to introduce disease resistance than conventional breeding.
Genetic Improvement through Biotechnology in Cereal Crops
A brief Power Point Presentation providing an overview about Genetic improvements through Biotechnology in Cereal Crops with precise and well known procedures and examples helping students about their academics on professional level in related fields.
Content for this compilation (Power Point Presentation) was collected from Different sources: Internet, books, social media, research papers, websites etc. and I definitely admire and acknowledge all these sources to provide such a helpful content.
This Presentation was prepared and presented during Bachelors degree in Agriculture (Session 2014-2018) with major subject of Plant Breeding and Genetics at "University College of Agriculture and Environmental Sciences, Baghdad-ul-Jadeed Campus, The Islamia University of Bahawalpur, Pakistan" as an Assignment on the topic of "Genetic Improvement Through Biotechnology in Cereal Crops".
I hope, content might be helpful on academic and professional level.
Best Regards
Muhammad Raza Ullah Tariq
Protein engineering
This document discusses protein engineering through directed evolution. It explains that directed evolution involves randomly recombining genes from protein libraries and screening mutant proteins for improved functions. This leaves beneficial changes up to chance but generates diversity. Rational design is also used to map important protein interactions to guide evolution. The document provides an example of using directed evolution to modify a cytochrome P450 enzyme to more efficiently convert alkanes to alcohols over multiple generations. However, it notes that expressing evolved proteins in vivo and requiring expensive cofactors limit the commercial potential of this approach.
ویرایش ژنوم Genome editing tools
Genome editing uses engineered nucleases to precisely modify DNA sequences and change the characteristics of cells and organisms. It involves using enzymes like CRISPR-Cas9 or zinc finger nucleases to create targeted double-stranded breaks in DNA, which are then repaired through non-homologous end joining or homology-directed repair. This allows genes to be knocked out, inserted, or altered. Genome editing has applications in research, biotechnology, agriculture, and potentially gene therapy. It works by harnessing the cell's own DNA repair mechanisms after targeted double strand breaks are introduced at specific genomic locations.
Pbr322 and puc8 plasmids
One of the first plasmids to be used in recombinant genetics was called pBR322. It is approximately 4300 bp in length and has two antibiotic resistance genes: Ap (Ampicillin) and Tc (Tetracycline). Bacteria cells that are successfully transformed with this plasmid are able to grow in the presence of both ampicillin and tetracycline antibiotics
Recent advancement recombinant dna technology
Recombinant DNA technology has led to many advancements in medicine and biotechnology. Key developments include:
1. The production of human insulin through recombinant DNA techniques, approved for use in 1982. This provided an effective treatment for diabetes and was the first commercial product of rDNA technology.
2. Vaccines produced using recombinant DNA techniques, such as vaccines for hepatitis B. This involves inserting the gene for the hepatitis B surface antigen protein into yeast which then produces the protein for the vaccine.
3. Production of other proteins through recombinant DNA like cytokines and interferons, which help modulate the immune system and can be used as treatments for various conditions. Overall, recombinant DNA technology has generated many commercially useful proteins and
Reverse Breeding
Reverse breeding is a novel plant breeding technique that allows the development of parental lines directly from any superior heterozygous plant. It involves suppressing meiotic recombination to produce gametes with whole parental chromosome sets, followed by doubling of haploids to generate parental lines. Two case studies demonstrate using RNAi to silence meiotic genes in Arabidopsis thaliana, producing parental lines that reconstitute the original hybrid when crossed. A second technique, marker-assisted reverse breeding, uses high-density SNP genotyping instead of gene silencing to select maize lines similar to original parents within one year. Reverse breeding techniques accelerate breeding and facilitate hybrid improvement without prior knowledge of parental lines.
Gene editing Dr. Abhinav Golla . MD pathology
Gene editing is a type of genetic engineering that allows scientists to precisely change an organism's DNA. It works by using engineered nucleases or "molecular scissors" to cut DNA at specific locations, which can then be repaired through natural cell processes or replaced with new DNA. The newest gene editing technique is called CRISPR-Cas9, which was adapted from a bacterial immune system and offers faster, cheaper, and more accurate editing compared to prior methods. CRISPR-Cas9 uses a small piece of RNA and an enzyme to target and cut specific DNA sequences.
Autobiography val
Valeria Rivera was born on December 17, 1992 in San Juan, Puerto Rico weighing 5 pounds and 10 ounces. She enjoys arts and crafts, knitting, bicycling, jogging, visiting beaches and swimming. She graduated high school with high honors in 2010 and is interested in studying veterinary medicine, having done volunteer work at a veterinary clinic.
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Genome Editing Introduction
Genome editing is a method of making specific changes to the DNA of a cell or organism. An enzyme scissors the DNA at a specific sequence, and when this is repaired by the cell, a change or ‘edit’ is made to the sequence.
https://www.creative-biolabs.com/gene-therapy/approaches-to-genome-editing.htm
Genome editing with engineered nucleases
Genome editing uses engineered nucleases to insert, replace or remove DNA from the genome. These nucleases create targeted double-strand breaks which are repaired through natural DNA repair processes, allowing for changes to the genome sequence. Three main engineered nuclease systems for genome editing are ZFNs, TALENs, and CRISPR-Cas9. CRISPR uses a guide RNA and Cas9 nuclease to make precise cuts at targeted DNA sequences for editing. It has advantages over ZFNs and TALENs in being cheaper, easier to design, and more efficient. Genome editing holds promise for applications in crops, medicine, and research.
Methods of screening
This document discusses various methods for screening recombinant clones, including:
1. Blue-white screening using X-gal to detect the presence or absence of insertions disrupting beta-galactosidase activity.
2. Insertional inactivation screening by inserting DNA fragments to disrupt antibiotic resistance or repressor genes.
3. Antibiotic sensitivity screening to detect circularized recombinant plasmids capable of conferring antibiotic resistance.
4. Auxotrophic yeast strain screening using yeast vectors containing selectable marker genes to complement host mutations.
5. Reporter gene assays using enzymes like luciferase, GFP, etc. fused to promoters to identify positive recombinant clones.
Genome editing
This presentation highlights the basics and application of genome editing strategies in plants, strategies to reduce off-target mutation, identification of mutant analysis etc.
P uc vectors
pUC vectors are plasmids derived from pBR322 that have a higher copy number of 500-600 copies per cell. They contain an ampicillin resistance gene for selection, as well as the lacZ' gene containing multiple cloning sites. When a gene of interest is inserted, it disrupts the lacZ' gene, allowing for blue-white screening on media containing IPTG and X-gal to identify recombinant colonies that appear white instead of blue. pUC vectors offer advantages over pBR322 such as high copy number and easy selection, though they cannot accommodate inserts larger than 15kb.
Zfn Compozr Mammalian Cle
Rapid and Precise Engineering of Mammalian Cells for GMP Manufacturing of Recombinant Proteins Pharmaceuticals
Insulin production by genetic engineering
This document outlines a lecture on microbial genetics. It discusses the genetic engineering processes used to produce human insulin and growth hormone (HGH) in bacteria. For insulin, the human gene is isolated and inserted into bacterial plasmid DNA using restriction enzymes. Transformed bacteria then produce insulin. A similar process is used for HGH, isolating its mRNA from human pituitary gland cells, generating cDNA, and inserting it into bacteria to produce HGH. Lastly, it notes other human hormones can be genetically engineered using these techniques.
Derivatives of pBR322
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.
Genome editing
genome editing is the emerging branch which includes the specific and precise engineering of genomic regions
Genome editing
Satrupa Das discusses using programmable nucleases like TALENs and CRISPR-Cas9 for genome editing to treat diseases. She describes a study where exon 44 was knocked back into the dystrophin gene of an iPSC line from a Duchenne muscular dystrophy patient using TALENs or Cas9. Over 90% of clones showed targeted integration of the donor template, and sequencing confirmed restoration of the reading frame without other mutations. Differentiated muscle cells from corrected clones expressed dystrophin, demonstrating therapeutic potential of genome editing for Duchenne muscular dystrophy.
Genome Editing Techniques by Kainat Ramzan
Genome technology has revolutionized biological science through techniques of Gene Editing in order to edit any organism's genome.MegNs and zinc-finger nucleases are commonly understood to be used, as is the effector's transcriptional activator-like nucleases. In CRISPR/Cas9, genetic alterations, and gene functionality have become a well-known tool for understanding gene targeting.
ZINC FINGER NUCLEASE TECHNOLOGY
Zinc finger nucleases (ZFNs) allow for highly targeted editing of the genome. ZFNs consist of a DNA-binding domain made of zinc finger proteins and a DNA-cleaving domain. The ZFN pair binds to a target site and creates a double-strand break, which the cell repairs through non-homologous end joining or homologous recombination, enabling gene knockouts or targeted changes. ZFNs work in many cell types and animal models, providing a more efficient alternative to traditional transgenic techniques. They have applications in functional genomics, cell line engineering, and animal model generation.
Crop genome editing using CRISPR
Genome editing uses tools like CRISPR/Cas9 to make precise, targeted changes to DNA in crop plants. The document discusses using CRISPR to edit the SlMlo1 gene in tomato, conferring resistance to powdery mildew. Researchers generated CRISPR/Cas9 plasmids targeting SlMlo1, transformed tomato plants, and identified lines with 48 bp deletions in the gene. Resistant T1 plants lacking the transgene were obtained, named the Tomelo variety. Genome editing provides a faster way to introduce disease resistance than conventional breeding.
Genetic Improvement through Biotechnology in Cereal Crops
A brief Power Point Presentation providing an overview about Genetic improvements through Biotechnology in Cereal Crops with precise and well known procedures and examples helping students about their academics on professional level in related fields.
Content for this compilation (Power Point Presentation) was collected from Different sources: Internet, books, social media, research papers, websites etc. and I definitely admire and acknowledge all these sources to provide such a helpful content.
This Presentation was prepared and presented during Bachelors degree in Agriculture (Session 2014-2018) with major subject of Plant Breeding and Genetics at "University College of Agriculture and Environmental Sciences, Baghdad-ul-Jadeed Campus, The Islamia University of Bahawalpur, Pakistan" as an Assignment on the topic of "Genetic Improvement Through Biotechnology in Cereal Crops".
I hope, content might be helpful on academic and professional level.
Best Regards
Muhammad Raza Ullah Tariq
Protein engineering
This document discusses protein engineering through directed evolution. It explains that directed evolution involves randomly recombining genes from protein libraries and screening mutant proteins for improved functions. This leaves beneficial changes up to chance but generates diversity. Rational design is also used to map important protein interactions to guide evolution. The document provides an example of using directed evolution to modify a cytochrome P450 enzyme to more efficiently convert alkanes to alcohols over multiple generations. However, it notes that expressing evolved proteins in vivo and requiring expensive cofactors limit the commercial potential of this approach.
ویرایش ژنوم Genome editing tools
Genome editing uses engineered nucleases to precisely modify DNA sequences and change the characteristics of cells and organisms. It involves using enzymes like CRISPR-Cas9 or zinc finger nucleases to create targeted double-stranded breaks in DNA, which are then repaired through non-homologous end joining or homology-directed repair. This allows genes to be knocked out, inserted, or altered. Genome editing has applications in research, biotechnology, agriculture, and potentially gene therapy. It works by harnessing the cell's own DNA repair mechanisms after targeted double strand breaks are introduced at specific genomic locations.
Pbr322 and puc8 plasmids
One of the first plasmids to be used in recombinant genetics was called pBR322. It is approximately 4300 bp in length and has two antibiotic resistance genes: Ap (Ampicillin) and Tc (Tetracycline). Bacteria cells that are successfully transformed with this plasmid are able to grow in the presence of both ampicillin and tetracycline antibiotics
Recent advancement recombinant dna technology
Recombinant DNA technology has led to many advancements in medicine and biotechnology. Key developments include:
1. The production of human insulin through recombinant DNA techniques, approved for use in 1982. This provided an effective treatment for diabetes and was the first commercial product of rDNA technology.
2. Vaccines produced using recombinant DNA techniques, such as vaccines for hepatitis B. This involves inserting the gene for the hepatitis B surface antigen protein into yeast which then produces the protein for the vaccine.
3. Production of other proteins through recombinant DNA like cytokines and interferons, which help modulate the immune system and can be used as treatments for various conditions. Overall, recombinant DNA technology has generated many commercially useful proteins and
Reverse Breeding
Reverse breeding is a novel plant breeding technique that allows the development of parental lines directly from any superior heterozygous plant. It involves suppressing meiotic recombination to produce gametes with whole parental chromosome sets, followed by doubling of haploids to generate parental lines. Two case studies demonstrate using RNAi to silence meiotic genes in Arabidopsis thaliana, producing parental lines that reconstitute the original hybrid when crossed. A second technique, marker-assisted reverse breeding, uses high-density SNP genotyping instead of gene silencing to select maize lines similar to original parents within one year. Reverse breeding techniques accelerate breeding and facilitate hybrid improvement without prior knowledge of parental lines.
Gene editing Dr. Abhinav Golla . MD pathology
Gene editing is a type of genetic engineering that allows scientists to precisely change an organism's DNA. It works by using engineered nucleases or "molecular scissors" to cut DNA at specific locations, which can then be repaired through natural cell processes or replaced with new DNA. The newest gene editing technique is called CRISPR-Cas9, which was adapted from a bacterial immune system and offers faster, cheaper, and more accurate editing compared to prior methods. CRISPR-Cas9 uses a small piece of RNA and an enzyme to target and cut specific DNA sequences.
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Genetic Improvement through Biotechnology in Cereal Crops
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Autobiography val
Valeria Rivera was born on December 17, 1992 in San Juan, Puerto Rico weighing 5 pounds and 10 ounces. She enjoys arts and crafts, knitting, bicycling, jogging, visiting beaches and swimming. She graduated high school with high honors in 2010 and is interested in studying veterinary medicine, having done volunteer work at a veterinary clinic.
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Proposal march 2012
This study aims to analyze conserved amino acids in GAPDH proteins from tropical plants Oxalis corniculata and Plectranthus amboinicus. GAPDH is important for energy production. Previous work cloned GAPDH genes from these plants and Myrtaceae psidium. Psidium showed no cloning and was eliminated. The current work will sequence the cloned GAPDH inserts and analyze conserved amino acids related to catalytic function through bioinformatics. This adds to knowledge of important plant genes and their evolution.
Valeria riverafinalpresentationdec2011v11 2
The author cloned GAPDH genes from Oxalis corniculata and Plectranthus amboinicus plants and conducted bioinformatics analyses. GAPDH is important for energy production. The genes were amplified by PCR and sequenced. Sequence analyses showed the cloned genes were GAPC from O. corniculata and GAPDH from P. amboinicus. Future work will include more detailed bioinformatics analyses and submitting the novel sequences to GenBank.
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This document summarizes a study that characterized the heme binding properties of glyceraldehyde-3-phosphate dehydrogenase (GAPDH). The key findings include:
1) GAPDH binds heme substoichiometrically, with one heme binding per GAPDH tetramer. The heme forms low-spin complexes with GAPDH that have distinct UV-visible absorption spectra depending on the heme redox state.
2) Kinetic analysis found heme binding to GAPDH is reversible and selective for heme structure. Heme binding affinity ranges from 19-390 nM depending on redox conditions.
3) Spectroscopic analysis indicates the heme in the GAPDH complex is bis-ligated by a histidine residue as the proximal
Crispr cas: A new tool of genome editing
The document summarizes a presentation on CRISPR cas9, a new genome editing tool. It discusses the history of CRISPR, how CRISPR functions in bacteria, the classification and components of CRISPR systems, and the mechanism of CRISPR cas9. It then covers applications of CRISPR cas9 in genome editing, databases of CRISPR sequences, case studies using the technology, and future directions of CRISPR research.
Recombi dna& medicine
Recombinant DNA technology allows genes to be isolated, modified, and re-expressed in other hosts. This document discusses several applications of recombinant DNA including producing human insulin through expressing the insulin gene in bacteria, using various cloning vectors like plasmids and phage to replicate genes of interest, and the potential use of HIV genes for gene delivery systems due to lentiviruses' ability to integrate into non-dividing cells. The document provides an overview of genetic engineering techniques and their medical applications.
Congreso de Biotecnología Arequipa Perú June 2011
This document summarizes research to bioengineer a new red fluorescent protein tag from a cyanobacteriochrome found in Thermosynechococcus elongatus. The researchers aim to develop a small, photostable red tag as an alternative to commonly used green and yellow fluorescent proteins from jellyfish. They use molecular biology techniques like PCR, site-directed mutagenesis, and transfection of E. coli and mammalian cells to express and purify the mutated protein, which they characterize through spectroscopy and microscopy. The goal is to create a useful new tool for cellular imaging applications.
Waksman Student Scholars Program Poster
The document describes an experiment to sequence the DNA of the duckweed plant Landoltia punctata and analyze the protein encoded. Researchers used PCR to amplify duckweed DNA inserted into bacteria. Restriction digest isolated the duckweed DNA, which was sequenced. Analysis identified the protein as Mitogen-Activated Protein Kinase (MAPK), which regulates cellular responses to stimuli and contains conserved domains. The 3D structure of this protein is similar to the human MAPK protein, indicating an evolutionary advantage across kingdoms.
Transplastomics
A transplastomic plant is a genetically modified plant in which the new genes have not been inserted in the nuclear DNA but in the DNA of the chloroplasts.
Powerpoint Final Paper RISE 2010
This experiment aims to isolate and sequence the GAPDH gene from the Cucumis sativus plant. GAPDH is an important enzyme that catalyzes a step in glycolysis. The experiment involves extracting DNA from C. sativus, amplifying the GAPDH gene via PCR, separating the DNA fragments via electrophoresis, purifying the DNA, and attempting to insert the gene into bacteria for replication and sequencing. However, the experiment was not completed because the bacteria needed for DNA purification did not grow, likely due to the presence of an antibiotic.
Engineering the plastid
The presentation describes the advantages of plastid transformation over 'conventional' nuclear transformation, hurdles to plastid transformation, its advantages. The presentation also covers some successful plastid engineering and its potential.
RNAi – Mechanism and Its Application In Crop Improvement
This document summarizes an RNAi presentation on crop improvement using RNA interference. The 3-sentence summary is:
RNA interference (RNAi) is a process of post-transcriptional gene silencing mediated by small RNA molecules. The presentation described the RNAi pathway and various applications of RNAi technology in crop improvement, including increasing nutrient levels, developing virus and pest resistance, and reducing anti-nutritional compounds. Several case studies were provided that demonstrated how RNAi has been used to successfully modify traits in different crops like maize, cotton, coffee, and banana.
written work Gapdh august 2009
This document summarizes a student research project that aims to sequence and analyze the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) genes from two plant species endemic to Puerto Rico: Cyperus alternifolius and Schefflera actinophylla. The student will transform and clone the GAPDH genes from these plants into E. coli, purify and sequence the cloned genes, and perform bioinformatics analysis to compare the sequences to other known GAPDH genes. The GAPDH gene codes for an important enzyme involved in cellular processes like glycolysis and apoptosis. While well-studied in model organisms, the GAPDH genes from many native Puerto Rican plants have yet to be sequenced.
Post genomic tools for genetic enhancement of germplasm
This document discusses how post-genomic tools like transcriptomics, proteomics, and metabolomics can be used for genetic enhancement of germplasm. It provides an introduction to each omics technique, examples of technologies used, and applications in understanding biological processes and identifying genes/proteins involved in traits. The conclusion states that omics expression analysis of germplasm will help characterize genome function and restore traits from wild varieties, aiding development of more sustainable crop varieties.
Molecular detection of food borne pathogens-presentation
This document provides an overview of molecular detection techniques used in food quality control. It discusses how chemistry alone cannot solve all detection problems and that molecular biology methods like PCR, RFLP, and sequencing are better alternatives as they are more accurate, rapid and cost-effective. It describes several common molecular detection methods and their applications in detecting food pathogens, adulterants, allergens and GM ingredients. The document emphasizes that molecular methods can identify microbes at the strain level and detect viable cells, but may not be able to find non-authorized GMOs due to lack of molecular information.
Genome Editing in Fruit Crops
This document discusses genome editing in fruit crops using CRISPR/Cas9 technology. It provides examples of using CRISPR to edit genes involved in fruit ripening, pigmentation, and flowering time regulation in strawberry, banana, apple, and kiwifruit. Specifically, it describes using CRISPR to increase beta-carotene levels in banana, induce early flowering in apple and pear, and generate dwarf kiwifruit plants. The document concludes that integrating biotechnology like CRISPR with conventional breeding is a promising strategy for fruit crop improvement.
Lab Differential Expression Differential gene expression provides th.pdf
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 .pdf
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.
JBEI June 2019 highlight slides
This document summarizes three studies related to plant biology and biochemistry. The first study engineered yeast and bacteria to produce bioactive phenolic compounds called hydroxycinnamoyl-L-amino acids (HAAs). The second study identified a gene involved in sphingolipid biosynthesis that modulates plasmodesmata structure and phloem transport in plants. The third study developed a web tool for predicting the functions of rice transcription factors based on gene expression patterns.
Bioinformatic analysis of the Proanthocyanidin Biosynthetic Pathway in Hawtho...
My master presentation of the project :
Bioinformatic analysis of the Proanthocyanidin Biosynthetic Pathway in Hawthorn (Crataegus spp.)
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RNAi – Mechanism and Its Application In Crop Improvement
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Lab Differential Expression Differential gene expression provides th.pdf
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Lab Differential Expression Differential gene expression provides .pdf
Lab Differential Expression Differential gene expression provides .pdf
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Synapses are specialized structures that transmit signals between neurons or from neurons to other cells. They convert electrical signals into chemical signals to allow communication between cells by releasing neurotransmitters in response to action potentials. Malfunctions in synaptic transmission can cause disorders like epilepsy, neuropathic pain, anxiety, depression, learning deficits, and dementia. The study focuses on how a calcium channel subunit called a25 that is targeted by anti-epileptic and pain drugs regulates synaptic transmission and structure.
The epilepsy paper 3
The study confirms associations between epilepsy in Australian Shepherd dogs and two locations on chromosomes 1 (CFA1) and 19 (CFA19) found in previous research. PCR-RFLP tests on 88 additional dogs showed different genotype frequencies between normal and affected dogs, supporting genes in these locations that may cause epilepsy. Sequencing of the DOK6 gene near associated SNPs on CFA1 found no differences between one affected and unaffected dog. The results support the hypothesis that mutations in these chromosomal regions contribute to epilepsy in Australian Shepherds.
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This study aimed to confirm previous findings linking canine chromosomes 1 and 19 to idiopathic epilepsy in Australian Shepherd dogs. PCR-RFLP tests of two SNPs were performed on 88 additional Australian Shepherds and genotypes were compared between affected and unaffected dogs via Chi-square analysis. Sequencing of the DOK6 gene did not reveal differences between one affected and unaffected dog. The results confirmed different genotype frequencies between normal and affected dogs for the SNPs, supporting the hypothesis that epilepsy-causing genes are located near the tested regions on chromosomes 1 and 19.
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This document summarizes research being done to identify genetic factors that cause epilepsy in Australian Shepherd dogs. The researchers are genotyping additional dogs to confirm associations between epilepsy and SNPs on chromosomes 1 and 19 found in previous work. They plan to sequence the DOK6 gene near one of the SNPs on chromosome 1 to look for mutations correlated with epilepsy. Current work includes genotyping 80 additional dogs and sequencing exons of the DOK6 gene, though one exon is still being analyzed. The goal is to identify the specific genetic cause of epilepsy in this breed to improve treatment and prevent transmission.
Lab 12 lv
This lab used an ELISA test to simulate the sharing of bodily fluids and determine which students were exposed to and transmitted a contagious disease. The ELISA test relies on antibodies to detect antigens produced during an immune response to exposure. The results showed students 1, 2, 8, and 10 were infected, while students 1 and 10 transmitted the disease. ELISA is commonly used in real world applications such as pregnancy tests, disease detection, and testing for food safety and GMOs.
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The document announces a nanotechnology workshop from October 7th to 9th organized by Ms. Ivonne Ferrer and Ms. Jesus Velazquez. The workshop will likely focus on nanotechnology topics over the three day period and is being run by the two individuals mentioned.
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The document discusses a university project studying epilepsy in Australian Shepherd dogs. It mentions the University of Minnesota St. Paul Campus LSSURP Project and names Valeria Rivera as the summer 2011 student and Ned Patterson as the mentor. The project focuses on epilepsy in Australian Shepherd dogs.
University of minnesota blog photos
The document discusses a university project studying epilepsy in Australian Shepherd dogs. It mentions the University of Minnesota St. Paul Campus LSSURP Project and names Valeria Rivera as the summer 2011 student and Ned Patterson as the mentor. The project focuses on epilepsy in Australian Shepherd dogs.
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The student reflects on their experience in the RISE program at UPR-Cayey. They took biology courses where they conducted laboratory work and wrote proposals. This helped them think like a scientist by asking questions and searching for information. They attended RISE seminars that introduced them to new scientific terms and potential research opportunities. This led the student to be selected for a summer research program at the University of Minnesota where they wrote annotated bibliographies and learned to communicate scientific findings. In their second semester, the RISE biomedical techniques course taught them new lab techniques to help achieve their career goals in the sciences.
Lab 12 e
This lab used an ELISA test to simulate the sharing of a contagious disease between students and determine who was infected and who transmitted the disease. The ELISA test relies on antibodies binding to antigens produced during an immune response to detect exposure to a disease in liquid samples. The results of the lab showed that students 1, 2, 8, and 10 were infected, while students 1 and 10 transmitted the disease to others.
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Beer's law states that absorbance is directly proportional to concentration, allowing this lab to use a spectrophotometer to calculate protein levels in milk samples. The students prepared 1:50 dilutions of evaporated and fresh milk with dye, incubated for 40 minutes, and measured absorbance at 595 wavelengths to generate a standard curve and determine that sample A, evaporated milk, had a higher protein concentration than the fresh milk.
Lab 10 e
Enzymes are proteins that act as catalysts and have specific shapes that determine their substrates and products. The experiment determined the optimum temperature of the enzyme cellobiase, which breaks down cellulose, to be 22°C. Samples of cellobiase were tested at 0°, 22°, and 37°C using a spectrophotometer to measure absorbance, finding that the enzyme works best at 22°C and its activity is minimized at higher temperatures due to denaturing.
Lab 9 e
During a lab experiment, students created a hydrogen sensor using palladium-coated polymer nanofibers produced through electrospinning and sputter coating, which was then tested in a chamber with controlled hydrogen and argon gas flows. The sensor worked effectively, with an increase in current detected as hydrogen levels rose, demonstrating an application of nanotechnology to increase surface area and catalytic reactivity for hydrogen detection.
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This document summarizes a lab assignment on bioinformatics. Students were asked to practice using bioinformatics tools like BLAST and databases from NCBI to analyze DNA sequences. For the assignment, students were given a scenario where they had to use these tools to identify which staff members at a research project were illegally using DNA from an endangered primate species. Students performed BLAST searches comparing DNA samples from staff members to a sequence from the endangered primate provided by another company, identifying that two staff members were implicated.
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- Step-by-step guide on deploying anomaly detection models on edge devices using ArgoCD.
5. Introduction to Apache Kafka and S3
- Explore Apache Kafka for real-time data streaming and Amazon S3 for scalable storage solutions.
6. Viewing Kafka Messages in the Data Lake
- Learn how to view and analyze Kafka messages stored in a data lake for better insights.
7. What is Prometheus?
- Get to know Prometheus, an open-source monitoring and alerting toolkit, and its application in monitoring edge devices.
8. Monitoring Application Metrics with Prometheus
- Detailed instructions on setting up Prometheus to monitor the performance and health of your anomaly detection system.
9. What is Camel K?
- Introduction to Camel K, a lightweight integration framework built on Apache Camel, designed for Kubernetes.
10. Configuring Camel K Integrations for Data Pipelines
- Learn how to configure Camel K for seamless data pipeline integrations in your anomaly detection workflow.
11. What is a Jupyter Notebook?
- Overview of Jupyter Notebooks, an open-source web application for creating and sharing documents with live code, equations, visualizations, and narrative text.
12. Jupyter Notebooks with Code Examples
- Hands-on examples and code snippets in Jupyter Notebooks to help you implement and test anomaly detection models.
How to Interpret Trends in the Kalyan Rajdhani Mix Chart.pdf
A Mix Chart displays historical data of numbers in a graphical or tabular form. The Kalyan Rajdhani Mix Chart specifically shows the results of a sequence of numbers over different periods.
HCL Notes und Domino Lizenzkostenreduzierung in der Welt von DLAU
Webinar Recording: https://www.panagenda.com/webinars/hcl-notes-und-domino-lizenzkostenreduzierung-in-der-welt-von-dlau/
DLAU und die Lizenzen nach dem CCB- und CCX-Modell sind für viele in der HCL-Community seit letztem Jahr ein heißes Thema. Als Notes- oder Domino-Kunde haben Sie vielleicht mit unerwartet hohen Benutzerzahlen und Lizenzgebühren zu kämpfen. Sie fragen sich vielleicht, wie diese neue Art der Lizenzierung funktioniert und welchen Nutzen sie Ihnen bringt. Vor allem wollen Sie sicherlich Ihr Budget einhalten und Kosten sparen, wo immer möglich. Das verstehen wir und wir möchten Ihnen dabei helfen!
Wir erklären Ihnen, wie Sie häufige Konfigurationsprobleme lösen können, die dazu führen können, dass mehr Benutzer gezählt werden als nötig, und wie Sie überflüssige oder ungenutzte Konten identifizieren und entfernen können, um Geld zu sparen. Es gibt auch einige Ansätze, die zu unnötigen Ausgaben führen können, z. B. wenn ein Personendokument anstelle eines Mail-Ins für geteilte Mailboxen verwendet wird. Wir zeigen Ihnen solche Fälle und deren Lösungen. Und natürlich erklären wir Ihnen das neue Lizenzmodell.
Nehmen Sie an diesem Webinar teil, bei dem HCL-Ambassador Marc Thomas und Gastredner Franz Walder Ihnen diese neue Welt näherbringen. Es vermittelt Ihnen die Tools und das Know-how, um den Überblick zu bewahren. Sie werden in der Lage sein, Ihre Kosten durch eine optimierte Domino-Konfiguration zu reduzieren und auch in Zukunft gering zu halten.
Diese Themen werden behandelt
- Reduzierung der Lizenzkosten durch Auffinden und Beheben von Fehlkonfigurationen und überflüssigen Konten
- Wie funktionieren CCB- und CCX-Lizenzen wirklich?
- Verstehen des DLAU-Tools und wie man es am besten nutzt
- Tipps für häufige Problembereiche, wie z. B. Team-Postfächer, Funktions-/Testbenutzer usw.
- Praxisbeispiele und Best Practices zum sofortigen Umsetzen
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Valeria proposalsat
- 1. Cloning and Bioinformatics Analyses of Novel Plant GAPDH Genes Valeria Rivera Dr. Michael Rubin RISE Program, Department of Biology, University of Puerto Rico at Cayey
- 2. Background Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) This gene codes for a protein that catalyzes a step of an important pathway, glycolysis, for energy production in carbohydrate metabolism. The human genome has an orthologous GAPDH gene. Techniques used: Polymerase Chain Reaction Electrophoresis Cloning and bacterial culture Plasmid Purification Bioinformatics analyses
- 3. Oxalis corniculata Sleeping Beauty http://www.ppws.vt.edu/scott/weed_id /oxast.htm
- 4. Plectranthus amboinicus Oregano brujo http://www.bitterrootrestoration.com/me dicinal-plants/plectranthus- .html
- 6. Significance Study of important genes involved in energy production in plants. Compare GAPDH genes from various plants to study their similarities and differences.
- 7. Problem Do GAPDH proteins have conserved amino acids related to the active site involved in catalytic function?
- 8. Hypothesis GAPDH proteins will have conserved amino acids related to the active site involved in catalytic function. GAPDH proteins will have different amino acids at other positions not related to the active site involved in catalytic function.
- 9. Specific Aims Design PCR Primers and Amplify GAPDH PCR Products Ligate PCR product into plasmid cloning vector pJET Transform Ligation Into competent E. coli Culture Bacteria Purify plasmid DNA, Digest with Restriction Endonucleases to Identify Cloned PCR Products Midi Prep – Large Scale Plasmid Purification Sequence the GAPDH Gene using Sanger Sequencing Bioinformatics Analyses
- 10. Sanger Sequencing The DNA template is copied. Terminators stop replication (fluorescent nucleotides). Fragments vary in length. Put samples on a plate in the sequencing machine using thin glass capillaries. Electrical field moves the negatively charged DNA to the positive pole through a gel matrix. A laser excites the labeled base creating a fluorescent color that is presented as a color peak.
- 11. Sanger Sequencing Each color peak represents a sequenced nucleotide. The sequence is determined by reading the different color patterns detected using a laser.
- 14. Acknowledgements RISE Program Lydia Cortés Dr. Michael Rubin
- 15. Cloning and Bioinformatics Analyses of Novel Plant GAPDH Genes Valeria Rivera Dr. Michael Rubin RISE Program, Department of Biology, University of Puerto Rico at Cayey
Editor's Notes
- Same between species
- All living organisms need energy. Energy is the ability to do work. Plants use light energy, carbon dioxide, and water to make sugar energy thorough photosynthesis releasing oxygen.
- Endonucleases adhere to the phosphodiester bond making the backbone of each helical strand (deoxyribose and ribose)
- Figure 14.13: Sanger sequencing. Following melting of DNA, PCR replication is performed with a small fraction of fluorescent dideoxynucleotides which terminate the polymerization process. Following replication, the strands, each of which terminates with a fluorophore, are separated and the readout gives the sequence.
- Did PCR, electrophoresis, and cloning bacterial culture. They ran an agarose gel that first shows the ladders or markers, then the negative controls followed by the positive controls. In the upper area of the gel are the cloning vectors that co-migrate. Then there are the 3 groups of plant samples. First is guava with no cloning since not enough DNA sample was extracted nor amplified as shown in the absence of PCR product band. In sleeping beauty, only two samples were cloned where as oregano brujo was successfully cloned.