Recombinant DNA Technology
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Recombinant DNA technology allows for the identification, isolation, modification, and re-expression of genes. The process involves isolating a gene of interest and inserting it into a bacterial plasmid using restriction enzymes and DNA ligase. This recombinant DNA is then inserted into bacteria via transformation. Key steps include using restriction endonucleases to cut DNA at specific recognition sequences, and bacteriophages' ability to infect and destroy bacteria unless their DNA is first destroyed by the host's restriction enzymes.
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rDna technology
rDNA technology involves deliberately manipulating the genetic makeup of cells or organisms in a controlled way. Its major goals are to form gene libraries, isolate and amplify genes of interest, and alter genetic makeup and gene expression patterns. The key steps are isolating DNA from cells, fragmenting the DNA with restriction enzymes, amplifying the gene of interest with PCR, and inserting the recombinant DNA into a host with enzymes.
Tools and tech. in Genetic Engineering and Restriction Enzymes by Kailash Son...
This document discusses genetic engineering tools and restriction enzymes. It describes how restriction enzymes are used to cut DNA at specific recognition sequences, producing either blunt or sticky ends. Some key restriction enzymes are described such as EcoRI, HindIII, and BamHI. The document also covers the discovery of restriction enzymes, their classification into Types I, II, and III, and their nomenclature based on the bacteria they were discovered in.
Unit 9 Manipulating Dna
1. Biotechnology relies on restriction enzymes that cut DNA at specific nucleotide sequences. Different enzymes cut DNA in different ways, leaving either blunt or sticky ends. Restriction maps show the lengths of DNA fragments cut by these enzymes.
2. The polymerase chain reaction (PCR) amplifies specific DNA sequences. It uses DNA polymerase to copy short DNA segments billions of times over, by cycling between high and low temperatures.
3. DNA fingerprinting identifies individuals by analyzing variations in noncoding DNA regions containing repeating sequences. The probability that any two individuals will have identical fingerprints across multiple regions is very small.
Restriction enzymes powerpoint (1)
Restriction enzymes are enzymes that break apart DNA into fragments or genes. They were originally created by bacteria as a defense mechanism to break down foreign DNA. There are three main types of restriction enzymes that recognize and cut DNA sequences in different ways. Restriction enzymes are commonly used to cut DNA into fragments for easier study and identification of genes, as well as recombining DNA molecules from different genomes to study gene expression and regulation.
Restriction of bacterial plasmid by Restriction Enzymes Practical
Restriction Enzymes are also known as Molecular scissors. Produce cohesive/sticky and blunt ends. Many restriction enzymes are present have their own particular Restriction sites (Palindromic Sequences)
Restriction Enzymes
Restriction enzymes are molecules that cut DNA at specific sequences. They were discovered in the 1970s and have become important tools in biotechnology. Restriction enzymes cut both strands of DNA in the same place and generate sticky or blunt ends. Daniel Nathans and colleagues were the first to isolate and characterize a restriction enzyme from Haemophilus influenzae that generates 11 fragments when cutting the SV40 virus DNA. Their work helped establish restriction enzymes as useful tools and earned them the 1978 Nobel Prize. Restriction enzymes are now widely used for cloning genes and studying DNA.
R-DNA technology and formation of r-DNA
Recombinant DNA technology involves joining DNA molecules from different species. This allows for new genetic combinations that can be inserted into host organisms. Recombinant DNA technology has many applications, including producing crops with desirable traits, manufacturing drugs like insulin, and conducting gene therapy. The process involves identifying a gene of interest, isolating it, inserting it into a vector like a plasmid, transforming the vector into a host cell, selecting transformed cells, and expressing the gene to multiply the product. This allows for large-scale production of proteins or genes of interest. While it provides benefits like substantial quantities of tailored products, concerns exist around its impacts on natural ecosystems and potential misuse.
Genetic engineering
This presentation contains b asic information regarding biotechnolgy and genetic engineering required for a food engineer and application of these to food sector.
Recommended
rDna technology
rDNA technology involves deliberately manipulating the genetic makeup of cells or organisms in a controlled way. Its major goals are to form gene libraries, isolate and amplify genes of interest, and alter genetic makeup and gene expression patterns. The key steps are isolating DNA from cells, fragmenting the DNA with restriction enzymes, amplifying the gene of interest with PCR, and inserting the recombinant DNA into a host with enzymes.
Tools and tech. in Genetic Engineering and Restriction Enzymes by Kailash Son...
This document discusses genetic engineering tools and restriction enzymes. It describes how restriction enzymes are used to cut DNA at specific recognition sequences, producing either blunt or sticky ends. Some key restriction enzymes are described such as EcoRI, HindIII, and BamHI. The document also covers the discovery of restriction enzymes, their classification into Types I, II, and III, and their nomenclature based on the bacteria they were discovered in.
Unit 9 Manipulating Dna
1. Biotechnology relies on restriction enzymes that cut DNA at specific nucleotide sequences. Different enzymes cut DNA in different ways, leaving either blunt or sticky ends. Restriction maps show the lengths of DNA fragments cut by these enzymes.
2. The polymerase chain reaction (PCR) amplifies specific DNA sequences. It uses DNA polymerase to copy short DNA segments billions of times over, by cycling between high and low temperatures.
3. DNA fingerprinting identifies individuals by analyzing variations in noncoding DNA regions containing repeating sequences. The probability that any two individuals will have identical fingerprints across multiple regions is very small.
Restriction enzymes powerpoint (1)
Restriction enzymes are enzymes that break apart DNA into fragments or genes. They were originally created by bacteria as a defense mechanism to break down foreign DNA. There are three main types of restriction enzymes that recognize and cut DNA sequences in different ways. Restriction enzymes are commonly used to cut DNA into fragments for easier study and identification of genes, as well as recombining DNA molecules from different genomes to study gene expression and regulation.
Restriction of bacterial plasmid by Restriction Enzymes Practical
Restriction Enzymes are also known as Molecular scissors. Produce cohesive/sticky and blunt ends. Many restriction enzymes are present have their own particular Restriction sites (Palindromic Sequences)
Restriction Enzymes
Restriction enzymes are molecules that cut DNA at specific sequences. They were discovered in the 1970s and have become important tools in biotechnology. Restriction enzymes cut both strands of DNA in the same place and generate sticky or blunt ends. Daniel Nathans and colleagues were the first to isolate and characterize a restriction enzyme from Haemophilus influenzae that generates 11 fragments when cutting the SV40 virus DNA. Their work helped establish restriction enzymes as useful tools and earned them the 1978 Nobel Prize. Restriction enzymes are now widely used for cloning genes and studying DNA.
R-DNA technology and formation of r-DNA
Recombinant DNA technology involves joining DNA molecules from different species. This allows for new genetic combinations that can be inserted into host organisms. Recombinant DNA technology has many applications, including producing crops with desirable traits, manufacturing drugs like insulin, and conducting gene therapy. The process involves identifying a gene of interest, isolating it, inserting it into a vector like a plasmid, transforming the vector into a host cell, selecting transformed cells, and expressing the gene to multiply the product. This allows for large-scale production of proteins or genes of interest. While it provides benefits like substantial quantities of tailored products, concerns exist around its impacts on natural ecosystems and potential misuse.
Genetic engineering
This presentation contains b asic information regarding biotechnolgy and genetic engineering required for a food engineer and application of these to food sector.
Restriction enzymes
Restriction enzymes are molecular scissors that cut DNA at specific recognition sequences. They play an important role in bacterial defense against invading viruses. There are four main types of restriction enzymes that differ in their composition, cofactors required, target sequences, and cleavage site positions. Type II enzymes are most commonly used in gene cloning and analysis. Restriction enzymes produce sticky or blunt ends that can be joined together through ligation. They have various applications including molecular cloning, DNA mapping, gene sequencing, restriction fragment length polymorphism analysis, pulsed field gel electrophoresis, and restriction enzyme-mediated integration.
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This document discusses recombinant DNA technology. It begins by defining recombinant DNA as DNA molecules formed by combining genetic material from multiple sources using genetic engineering techniques. The key steps involved are isolating genetic material, restriction enzyme digestion, amplification via PCR, ligating DNA molecules, inserting the recombinant DNA into a host, and isolating recombinant cells. The document then discusses each step in more detail and provides examples of applications like insulin production and Bt cotton. It concludes by noting some limitations like potential environmental impacts and vulnerability of cloned populations.
Digestion in gene cloning
Restriction enzyme digestion is a common technique used in gene cloning to prepare DNA fragments for insertion into a vector. Restriction enzymes recognize specific sequences in DNA and cleave the bonds between nucleotides at that site. The amount of restriction enzyme needed in microliters to completely digest one microgram of substrate DNA in one hour at the optimal temperature is defined as one unit of restriction enzyme activity. Restriction digestion is performed as part of gene cloning to isolate the desired DNA fragment, which is then inserted into a vector for cloning.
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Genetic engineering and Recombinant DNA
Genetic engineering involves altering the DNA of living organisms using biotechnology. It includes techniques like changing single DNA base pairs, deleting or adding genes, or combining DNA from different species. Recombinant DNA technology is used to create recombinant DNA molecules by manipulating DNA in vitro and introducing them into host organisms. This allows bacteria to be engineered to produce human insulin through inserting the human insulin gene into bacterial plasmids. Genomic libraries can be created by ligating fragmented genomic or cDNA into plasmid vectors to transform bacteria and clone the entire genome.
Restriction enzyme
Mata kuliah tentang enzim restriksi.Cari lebih banyak lagi di: http://muhammadhabibielecture.blogspot.com/2015/02/materi-kuliah-semester-4.html
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RESTRICTION ENZYMES
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Biotechnological tools used for diagnostic
This document discusses several biotechnological tools used for diagnostics:
1. DNA isolation, restriction enzyme digestion, polymerase chain reaction (PCR), gel electrophoresis, and DNA sequencing. PCR is described as artificially multiplying genetic material using primers and DNA polymerase. DNA sequencing methods like Sanger sequencing and Maxam-Gilbert are also outlined. The document concludes by briefly discussing gene cloning techniques like recombinant DNA technology and PCR for preparing copies of DNA fragments.
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Restriction endonucleases are enzymes found in bacteria that recognize specific nucleotide sequences in DNA and cut the DNA at those sites. This acts as a defense mechanism for bacteria by destroying foreign DNA, such as that from viruses. There are several types of restriction endonucleases, but type II are most useful for genetic engineering as they cut DNA at predictable, site-specific locations. The resulting DNA fragments can be joined back together via DNA ligases. Restriction endonucleases have four- to six-letter recognition sequences that are palindromic, and they produce sticky or blunt ends depending on whether cuts are offset or at the same position.
Biotech labs - restriction digest and transformation
This document summarizes 3 labs:
1. Restriction digest of lambda DNA - Use enzymes to cut lambda DNA into fragments, then analyze the fragments using gel electrophoresis. 5 fragments would be produced and the smallest would be largest.
2. Bacterial transformation - Insert GFP gene from jellyfish into bacterial DNA on a plasmid, then insert plasmid into bacteria. The bacteria could then express GFP.
3. DNA fingerprinting - Use restriction enzymes to cut DNA samples, separate fragments by size using gel electrophoresis, then compare fragment patterns to identify related samples or find matches for crime scene DNA.
389 b68b5 72c0-4364-a687-c85bfd929865
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This document presents on the role of active DNA demethylation in plant-virus interactions. It discusses geminiviruses, DNA methylation, and the general objective. It describes methods that will be used such as RT qPCR, co-immunoprecipitation, and Western blot analysis. Results are presented in Figures 1, 2, and 4. The discussion cites several references and findings about DME expression leading to hypermethylation and downregulation of genes. It concludes that genetic testing can help identify which drug works best against a specific virus to avoid pathogenesis.
Gene silencing
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Virus induced gene silencing
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Cloning and characterization of full length candidate genes
This document discusses cloning and characterization of candidate genes for physiological traits. It defines candidate genes as genes of known biological function involved in trait development or expression. It describes how candidate genes are identified and developed, and how genes are then cloned using restriction enzymes and vectors to replicate the target gene. Gene characterization is described as determining the expression of heritable traits through molecular analysis and genotyping. Three case studies demonstrate identifying drought tolerance genes in cassava, a dehydrin gene conferring stress tolerance in bajra, and isolating a phytocystatin gene for pest and disease resistance in turmeric.
Allozymes and isozymes
The document discusses allozymes and isozymes, which are genetic markers used in ichthyotaxonomy to study phylogenetic relationships between fish species. Allozymes represent enzymes catalyzed by different alleles of the same gene, while isozymes are enzymes from different genes that catalyze the same reaction. Allozyme electrophoresis is used to detect genetic variation at the enzyme level by identifying different allozyme forms. While allozymes and isozymes provide useful information and are easy to use, they have limitations as only a small fraction of enzyme loci are polymorphic and a limited number can be analyzed.
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Restriction endonucleases are enzymes produced by bacteria and archaea that recognize specific DNA sequences and cleave the sugar-phosphate backbone at or near the recognition site. In 1968, Werner Arber showed that an E. coli cell extract restricted fd phage DNA, demonstrating restriction in vitro. In 1970, Hamilton Smith isolated the first restriction enzyme, HindII, from Haemophilus influenzae. Daniel Nathans later showed specific cleavage of SV40 DNA by HindII. Restriction enzymes have been widely used in recombinant DNA technology and classified into four main types based on composition, cofactor requirements, target sequence, and cleavage position.
Recombinant DNA Technology
Recombinant DNA technology involves creating copies of DNA or genes by inserting them into vectors for introduction into host organisms. The key steps are: 1) isolating the gene of interest, 2) inserting it into a vector using restriction enzymes and DNA ligase, 3) transforming the recombinant DNA into a host cell, and 4) allowing the host to multiply and express the inserted gene. Common applications include producing recombinant human insulin, vaccines, and engineering crops for traits like herbicide or insect resistance.
rDNA Technology
Recombinant DNA technology involves combining DNA molecules from two different sources into one molecule by cutting and joining DNA strands. It includes three main tools - restriction enzymes, vectors, and host organisms. Restriction enzymes cut DNA at specific sites, vectors carry recombinant DNA, and host organisms allow the expression of inserted genes. The process involves isolating DNA, cutting with restriction enzymes, joining DNA segments by ligation, inserting into a host, and amplifying the target gene product. It has many applications in research, medicine, and industry, but also poses limitations such as environmental impacts and creating disease-causing organisms.
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Restriction enzymes
Restriction enzymes are molecular scissors that cut DNA at specific recognition sequences. They play an important role in bacterial defense against invading viruses. There are four main types of restriction enzymes that differ in their composition, cofactors required, target sequences, and cleavage site positions. Type II enzymes are most commonly used in gene cloning and analysis. Restriction enzymes produce sticky or blunt ends that can be joined together through ligation. They have various applications including molecular cloning, DNA mapping, gene sequencing, restriction fragment length polymorphism analysis, pulsed field gel electrophoresis, and restriction enzyme-mediated integration.
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This document discusses recombinant DNA technology. It begins by defining recombinant DNA as DNA molecules formed by combining genetic material from multiple sources using genetic engineering techniques. The key steps involved are isolating genetic material, restriction enzyme digestion, amplification via PCR, ligating DNA molecules, inserting the recombinant DNA into a host, and isolating recombinant cells. The document then discusses each step in more detail and provides examples of applications like insulin production and Bt cotton. It concludes by noting some limitations like potential environmental impacts and vulnerability of cloned populations.
Digestion in gene cloning
Restriction enzyme digestion is a common technique used in gene cloning to prepare DNA fragments for insertion into a vector. Restriction enzymes recognize specific sequences in DNA and cleave the bonds between nucleotides at that site. The amount of restriction enzyme needed in microliters to completely digest one microgram of substrate DNA in one hour at the optimal temperature is defined as one unit of restriction enzyme activity. Restriction digestion is performed as part of gene cloning to isolate the desired DNA fragment, which is then inserted into a vector for cloning.
about genetic modified orgenism
This document provides information on genetically modified organisms (GMOs), including their production process and applications in crop improvement. It defines GMOs and discusses common genetic modification techniques like recombinant DNA technology, genetic engineering, and gene splicing. The key steps in GMO production are isolating the gene of interest, inserting it into a vector, transforming the host organism, and selecting transformed offspring. Common transformation methods include Agrobacterium-mediated transformation and particle bombardment. The document also outlines applications of GMOs in increasing crop yields and resistance to pests and diseases.
Genetic engineering and Recombinant DNA
Genetic engineering involves altering the DNA of living organisms using biotechnology. It includes techniques like changing single DNA base pairs, deleting or adding genes, or combining DNA from different species. Recombinant DNA technology is used to create recombinant DNA molecules by manipulating DNA in vitro and introducing them into host organisms. This allows bacteria to be engineered to produce human insulin through inserting the human insulin gene into bacterial plasmids. Genomic libraries can be created by ligating fragmented genomic or cDNA into plasmid vectors to transform bacteria and clone the entire genome.
Restriction enzyme
Mata kuliah tentang enzim restriksi.Cari lebih banyak lagi di: http://muhammadhabibielecture.blogspot.com/2015/02/materi-kuliah-semester-4.html
RNA interference (RNAi)
RNA interference is a mechanism by which small RNA molecules like microRNA and siRNA regulate gene expression by binding to mRNA and preventing protein production. This mechanism is important for immunity and controlling gene expression. It was discovered in 1998 by Fire and Mello through experiments in C. elegans showing that double-stranded RNA, but not single stranded, could efficiently silence genes. RNAi has many applications in biotechnology, functional genomics, and medicine.
RESTRICTION ENZYMES
BRIEFLY EXPLAINED PPT ABOUT RESTRICTION ENZYMES, THEIR WORKING SITES, TYPES, ARTIFICIALLY GENERATED RESTRICTION ENZYMES, THEIR MECHANISM OF ACTION, TYPES OF CUTS THEY MAKE, THEIR NOMENCLATURE ETC.
140604 p03
This document provides an overview of current and future RNAi applications in plants. It discusses using RNAi to engineer resistance in plants against various pathogens like nematodes, insects, fungi, viruses, and parasitic weeds. It also discusses using RNAi to manipulate plant metabolism to improve industrial traits like fiber quality and oil content, or nutritional value by increasing carotenoids, amylose content, and reducing gluten proteins harmful to celiac disease patients. Specific examples demonstrating successful RNAi-mediated resistance or trait improvement in crops are provided.
Biotechnological tools used for diagnostic
This document discusses several biotechnological tools used for diagnostics:
1. DNA isolation, restriction enzyme digestion, polymerase chain reaction (PCR), gel electrophoresis, and DNA sequencing. PCR is described as artificially multiplying genetic material using primers and DNA polymerase. DNA sequencing methods like Sanger sequencing and Maxam-Gilbert are also outlined. The document concludes by briefly discussing gene cloning techniques like recombinant DNA technology and PCR for preparing copies of DNA fragments.
RESTRICTION ENDONUCLEASES & DNA LIGASES SMG
Restriction endonucleases are enzymes found in bacteria that recognize specific nucleotide sequences in DNA and cut the DNA at those sites. This acts as a defense mechanism for bacteria by destroying foreign DNA, such as that from viruses. There are several types of restriction endonucleases, but type II are most useful for genetic engineering as they cut DNA at predictable, site-specific locations. The resulting DNA fragments can be joined back together via DNA ligases. Restriction endonucleases have four- to six-letter recognition sequences that are palindromic, and they produce sticky or blunt ends depending on whether cuts are offset or at the same position.
Biotech labs - restriction digest and transformation
This document summarizes 3 labs:
1. Restriction digest of lambda DNA - Use enzymes to cut lambda DNA into fragments, then analyze the fragments using gel electrophoresis. 5 fragments would be produced and the smallest would be largest.
2. Bacterial transformation - Insert GFP gene from jellyfish into bacterial DNA on a plasmid, then insert plasmid into bacteria. The bacteria could then express GFP.
3. DNA fingerprinting - Use restriction enzymes to cut DNA samples, separate fragments by size using gel electrophoresis, then compare fragment patterns to identify related samples or find matches for crime scene DNA.
389 b68b5 72c0-4364-a687-c85bfd929865
This document presents on the role of active DNA demethylation in plant-virus interactions. It discusses geminiviruses, DNA methylation, and the general objective. It describes methods that will be used such as RT qPCR, co-immunoprecipitation, and Western blot analysis. Results are presented in Figures 1, 2, and 4. The discussion cites several references and findings about DME expression leading to hypermethylation and downregulation of genes. It concludes that genetic testing can help identify which drug works best against a specific virus to avoid pathogenesis.
7539 a744 19a8-42c3-a1e8-e3a20045a125
This document presents on the role of active DNA demethylation in plant-virus interactions. It discusses geminiviruses, DNA methylation, and the general objective. It describes methods that will be used such as RT qPCR, co-immunoprecipitation, and Western blot analysis. Results are presented in Figures 1, 2, and 4. The discussion cites several references and findings about DME expression leading to hypermethylation and downregulation of genes. It concludes that genetic testing can help identify which drug works best against a specific virus to avoid pathogenesis.
Gene silencing
Gene silencing is an epigenetic process that reduces or eliminates the production of a protein from its corresponding gene. There are two main types: transcriptional gene silencing via histone modifications, and post-transcriptional gene silencing by destroying or blocking mRNA. Methods to induce gene silencing include antisense oligonucleotides, ribozymes, and RNA interference using short interfering RNA or microRNA. RNAi leads to sequence-specific degradation of mRNA through the RNA-induced silencing complex. Gene silencing has applications in studying gene function, functional genomics, and medicine, with the goal of downregulating gene expression.
RNA interference technologies to control pests and pathogens - Steve Whyard -...
10-12 April 2019: The OECD Conference on RNAi based pesticides provided an overview on the current status and future possibilities for the regulation of externally applied dsRNA-based products that are proposed for use as pesticides. The event facilitated exchanges between policy makers, academia, industry on their implications in health, environment, and regulation.
Virus induced gene silencing
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Cloning and characterization of full length candidate genes
This document discusses cloning and characterization of candidate genes for physiological traits. It defines candidate genes as genes of known biological function involved in trait development or expression. It describes how candidate genes are identified and developed, and how genes are then cloned using restriction enzymes and vectors to replicate the target gene. Gene characterization is described as determining the expression of heritable traits through molecular analysis and genotyping. Three case studies demonstrate identifying drought tolerance genes in cassava, a dehydrin gene conferring stress tolerance in bajra, and isolating a phytocystatin gene for pest and disease resistance in turmeric.
Allozymes and isozymes
The document discusses allozymes and isozymes, which are genetic markers used in ichthyotaxonomy to study phylogenetic relationships between fish species. Allozymes represent enzymes catalyzed by different alleles of the same gene, while isozymes are enzymes from different genes that catalyze the same reaction. Allozyme electrophoresis is used to detect genetic variation at the enzyme level by identifying different allozyme forms. While allozymes and isozymes provide useful information and are easy to use, they have limitations as only a small fraction of enzyme loci are polymorphic and a limited number can be analyzed.
L9. restriction endonucleases
Restriction endonucleases are enzymes produced by bacteria and archaea that recognize specific DNA sequences and cleave the sugar-phosphate backbone at or near the recognition site. In 1968, Werner Arber showed that an E. coli cell extract restricted fd phage DNA, demonstrating restriction in vitro. In 1970, Hamilton Smith isolated the first restriction enzyme, HindII, from Haemophilus influenzae. Daniel Nathans later showed specific cleavage of SV40 DNA by HindII. Restriction enzymes have been widely used in recombinant DNA technology and classified into four main types based on composition, cofactor requirements, target sequence, and cleavage position.
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Similar to Recombinant DNA Technology
Recombinant DNA Technology
Recombinant DNA technology involves creating copies of DNA or genes by inserting them into vectors for introduction into host organisms. The key steps are: 1) isolating the gene of interest, 2) inserting it into a vector using restriction enzymes and DNA ligase, 3) transforming the recombinant DNA into a host cell, and 4) allowing the host to multiply and express the inserted gene. Common applications include producing recombinant human insulin, vaccines, and engineering crops for traits like herbicide or insect resistance.
rDNA Technology
Recombinant DNA technology involves combining DNA molecules from two different sources into one molecule by cutting and joining DNA strands. It includes three main tools - restriction enzymes, vectors, and host organisms. Restriction enzymes cut DNA at specific sites, vectors carry recombinant DNA, and host organisms allow the expression of inserted genes. The process involves isolating DNA, cutting with restriction enzymes, joining DNA segments by ligation, inserting into a host, and amplifying the target gene product. It has many applications in research, medicine, and industry, but also poses limitations such as environmental impacts and creating disease-causing organisms.
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Recombinant DNA technology involves combining DNA from different sources and inserting it into a host organism. Key tools used include restriction enzymes to cut DNA, vectors to carry inserted DNA, and host organisms like bacteria. The process involves selecting a DNA insert, integrating it into a vector, transforming a host, and having the host express the inserted DNA. Applications include producing human insulin, vaccines, herbicide-resistant crops, and diagnosing HIV.
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This document provides an overview of techniques for exploring genes, including DNA purification, restriction enzymes, recombinant DNA technology, molecular cloning using vectors, DNA libraries, blotting techniques, DNA sequencing, PCR, and gene expression studies. Key concepts covered include the types of DNA, plasmid purification, DNA conformations, nucleic acid quantitation, gel electrophoresis, restriction enzyme recognition sites and nomenclature, recombinant DNA technology tools, molecular cloning steps and components of cloning vectors. Applications of these techniques such as genetically modified organisms, recombinant proteins, gene therapy, and CRISPR are also discussed.
Genetic engineering
Genetic Engineering, also called as recombinant DNA technology, involves the group of techniques used to cut up and join together genetic material, especially DNA from different biological species, and to introduce the resulting hybrid DNA into an organism in order to form new combinations of heritable genetic material. This slide will illustrate the basic concepts and steps involved in Genetic Engineering.
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Gene transplantation is a method to select genes from cells using recombinant DNA techniques, manipulate the genes, and introduce them into other organisms to provide benefits. The process involves isolating the gene of interest using restriction endonucleases, producing hybrid DNA by combining the gene with a vector like a virus or plasmid, and incorporating the recombinant DNA into a recipient cell where it can be cloned. Restriction endonucleases recognize specific DNA sequences and cleave DNA into fragments that can be used in gene transplantation by separating fragments on gels according to molecular weight.
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This document provides information about the course "Introduction to Biotechnology" and the lesson on "Recombinant DNA Technology". It defines recombinant DNA as DNA formed by combining genetic material from different sources. It describes the steps to create recombinant DNA which include isolating DNA fragments, cutting them with restriction enzymes, joining them together with ligase, and inserting them into a host cell. It provides examples of using recombinant DNA technology to produce insulin in bacteria.
F tools of genetic engineering
Tools of genetic engineering include restriction enzymes, DNA ligase, DNA polymerase, and cloning vectors. Restriction enzymes cut DNA at specific recognition sequences, leaving sticky or blunt ends. DNA ligase joins DNA fragments by sealing nicks in DNA strands. DNA polymerase synthesizes new strands of DNA using existing strands as templates. Cloning vectors, such as plasmids, are used to introduce foreign DNA into host cells for amplification. Key steps in gene cloning are isolation of the gene, restriction digestion, ligation into a vector, transformation into host cells, and screening for the recombinant DNA.
Gene Cloning.ppt
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA.
DNA cloning is a molecular biology technique that makes many identical copies of a piece of DNA, such as a gene.
In a typical cloning experiment, a target gene is inserted into a circular piece of DNA called a plasmid.
The plasmid is introduced into bacteria via a process called transformation, and bacteria carrying the plasmid are selected using antibiotics.
Bacteria with the correct plasmid are used to make more plasmid DNA or, in some cases, induced to express the gene and make protein.
BASIC STEPS OF GENE CLONING are :
1. Cutting and pasting DNA and Vectors
2. Bacterial transformation and selection
3.Grow up lots of plasmid-carrying bacteria and use them as "factories" to make the protein. Harvest the protein from the bacteria and purify it
Basic Requirements for Gene Cloning:
Cloning Vectors
Isolation Procedure for DNA fragment and Vectors
Appropriate Enzyme for Purified DNA manipulation
Host Cells
Agar with Antibiotics
Vectors for Gene Cloning: Plasmid and Bacteriophage :
Characteristic of a Vector for Gene cloning
It must be able to replicate with in the host cells
It needs to be small , Ideally less than 10 kb in size
Two kind of DNA molecule that satisfy the above criteria can be found in bacterial cells, namely Plasmid and Bacteriophage chromosomes
Plasmid:Is a circular molecule of DNA that lead un independent existence in the bacteria cells.
Bacteriophages, or phages are viruses that specifically
infect bacteria.
Characteristic of a Vector for Gene cloning
It must be able to replicate with in the host cells
It needs to be small , Ideally less than 10 kb in size
Two kind of DNA molecule that satisfy the above criteria can be found in bacterial cells, namely Plasmid and Bacteriophage chromosomes
Plasmid:Is a circular molecule of DNA that lead un independent existence in the bacteria cells.
Bacteriophages, or phages are viruses that specifically
infect bacteria.
Characteristic of a Vector for Gene cloning
It must be able to replicate with in the host cells
It needs to be small , Ideally less than 10 kb in size
Two kind of DNA molecule that satisfy the above criteria can be found in bacterial cells, namely Plasmid and Bacteriophage chromosomes
Plasmid:Is a circular molecule of DNA that lead un independent existence in the bacteria cells.
Bacteriophages, or phages are viruses that specifically
infect bacteria.
The procedure for total DNA preparation from a culture of bacterial cells can be divided into four stages (Figure 3.1):
A culture of bacteria is grown and then harvested.
The cells are broken open to release their contents.
This cell extract is treated to remove all components except the DNA.
The resulting DNA solution is concentrated
Treatment with EDTA and lysozyme is carried out in the presence
of sucrose, which prevents the cells from bursting immediately.
Construction of rDNA molecules and bacterial transformation
Recombinant DNA technology involves introducing genes of interest into the genome of an organism using genetic engineering techniques. This is done by cutting DNA fragments using restriction enzymes, joining the fragments to a vector plasmid, and introducing the recombinant DNA or rDNA into a host cell. The host cell then replicates and produces multiple copies of the rDNA. This allows genes from one species to be introduced into another, such as joining human DNA to E. coli bacteria. The rDNA can then be used to produce beneficial proteins and genetically modified organisms.
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This document discusses the key enzymes involved in recombinant DNA technology. It describes how restriction enzymes cut DNA at specific recognition sites, and DNA ligases join cut DNA fragments back together. The document outlines the process of recombinant DNA technology, including generating DNA fragments, inserting them into cloning vectors, introducing the vectors into host cells, and expressing the gene of interest. It provides details on various restriction enzymes and DNA-modifying enzymes used in genetic engineering applications.
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This document is a biology investigatory project submitted by Arajit Kumar Pati on the topic of genetic engineering. It provides an overview of genetic engineering and recombinant DNA technology. It describes various techniques used in genetic engineering like DNA cloning, restriction digestion, use of vectors, recombinant DNA formation, insertion of genes into hosts, and production of gene products. It discusses the wide applications of genetic engineering in fields like medicine, agriculture, and industry. It concludes that genetic engineering has revolutionized our understanding of inheritance and molecular biology.
Assignment on Recombinant DNA Technology and Gene Therapy
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Recombinant DNA.1.ppt
Recombinant DNA technology involves taking genes from one organism and introducing them into another. The key steps are:
1. Isolating the desired DNA fragment and cutting it with restriction enzymes.
2. Inserting the fragment into a cloning vector, such as a plasmid, by ligating the DNA pieces together.
3. Introducing the recombinant DNA into a host organism, usually bacteria, through a process called transformation. The host then replicates and expresses the inserted gene.
Lecture 1 molecular tech. rdt 11 50-2020
This document discusses recombinant DNA technology (RDT), including its key steps and applications. RDT involves manipulating DNA sequences to produce chimeric DNA molecules using genetic engineering techniques. Restriction endonucleases are used to cut DNA at specific sequences. Vectors like plasmids, bacteriophages and cosmids are used to insert and replicate foreign DNA. RDT has applications like producing therapeutic proteins, vaccines, gene therapy and understanding disease mechanisms. It has revolutionized biology and medicine.
Study of cloning vectors and recombinant dna technology
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29112019 Sahri Yanti DNA Recombinan Technology
DNA recombinant technology involves manipulating genes in organisms. It uses restriction enzymes to cut DNA at specific sites, vectors to carry foreign DNA, and host organisms for transformation. The key steps are:
1) Isolating the gene of interest from a library or by synthesis
2) Inserting the gene into a vector like a plasmid using enzymes and ligation
3) Transforming the vector into a host organism through physical or chemical methods
4) Selecting transformed host cells that contain the recombinant DNA
5) Analyzing the new genes and proteins produced by the host.
Lecture 1 molecular tech. RDT By Dr Vishnu Kumar Professor, Biochemistry
This document discusses recombinant DNA technology, including defining key terms like restriction endonucleases, sticky ends, blunt ends, cloning, vectors, gene therapy, and DNA libraries. It describes how restriction enzymes cut DNA, how sticky and blunt ends are joined, the role of plasmids, bacteriophages and cosmids as vectors, and applications of recombinant DNA technology such as producing insulin to treat diabetes.
RECOMBINANT DNA TECHNOLOGY PRESENTATIONS
Recombinant DNA technology involves manipulating and combining DNA from different species using laboratory techniques. The resulting recombinant DNA can be used to produce large quantities of proteins encoded by genes of interest. Key steps include isolating the desired DNA and vector, combining them to form recombinant DNA, introducing this into a host cell, and causing the host cell to multiply and express the gene to produce the desired protein product. Various tools are used, including restriction enzymes to cut DNA, DNA ligase to join DNA, and polymerases for DNA replication. Common gene transfer techniques include transformation, transduction, electroporation, conjugation, and direct DNA transfer.
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Recombinant DNA Technology
- 3. • Recombinant DNA is a tool in understanding the structure, function , and regulation of genes and their products.
- 4. The objective of Recombinant DNA technology include ; _Identifying genes. _ Isolating genes . _ Modifying genes . _ Re-expressing genes in other hosts or organisms .
- 5. Cloning Steps : • Step 1 : DNA Isolation _ Isolation of your gene of Interest . .Step 2 : Recombinant DNA _ Insertion of foreign DNA into bacterial plasmid using restriction enzymes and DNA Ligase . . Step 3 : Transformation Insertion of recombinant DNA into bacteria by making bacteria component . . Step 4 : Production of recombinant DNA .
- 7. Enzymes • Enzymes are proteins - biological catalysts → help drive biochemical reactions. • Enzymes names end with an ase (e.g.., endonuclease ) _ Bacteria have evolved a class of enzymes that destroy foreign DNA (e.g.. Virus DNA). Protect bacteria from bacteriophages (Virus). _ Bacteriophages cannot multiply if their DNA is destroyed by host .
- 8. Restriction Endonucleases • Restriction endonucleases RESTRICT viruses - viral genome is destroyed upon entry. • Restriction endonucleases = Restriction enzymes - Endo (inside), nuclease (cuts nucleic acid) • Restriction endonucleases recognizes a short and specific DNA sequence and cuts it from insides. • The specific DNA sequence is called recognition sequence.
- 9. Bacteriophage Life Graph • Animation of the Lytic Life cycle of a Bacteriophage
- 10. Nomenclature • Smith and Nathans (1973) proposed enzymes naming scheme . - three-letter acronym for each enzyme derived from the source organism - Frist letter fro genus - Next two letters represent species - Additional latter or number represent the strain or serotypes • For example: The enzyme Hindll was isolated from Haemophilus influenzae serotyped.