Recombinant DNA technology allows for the manipulation of genes from different species in the laboratory. The process involves isolating the gene of interest, inserting it into a vector, and introducing the vector into a host cell. This allows the gene to be expressed, producing its protein product in large quantities. Key tools that enable recombinant DNA techniques are restriction enzymes, DNA ligase, bacterial plasmids as vectors, and methods for introducing DNA into host cells like transformation, transduction, and microinjection. Recombinant DNA technology has applications in producing foods, antibiotics, enzymes and more through genetic engineering of microbes, plants and animals.
This document describes various methods for transferring genes into organisms. Biological methods include using viruses like cauliflower mosaic virus (CaMV) to transfer genes into plants. Physical methods include electroporation, which uses electric pulses to create pores in cell membranes through which DNA can enter. Liposomes and direct methods like microinjection and particle bombardment can also be used to directly transfer DNA. Chemical methods involve using compounds like polyethylene glycol (PEG) to destabilize cell membranes and allow DNA uptake. While physical methods can target single cells, they may damage cells. Biological methods using vectors are often more efficient but less controlled. Overall the document provides an overview of the key gene transfer techniques.
introduction
What is virus
What is virus resistance plant
History
Gene use for develop virus resistance plant
Coat protein gene
cDNA of satellite RNA
Defective viral genome
Antisense RNA approach and
Ribozyme – mediated protection
conclusion
References
The document summarizes the ligase chain reaction (LCR) technique. LCR amplifies DNA sequences using four probes and ligase and polymerase enzymes instead of amplifying DNA through nucleotide polymerization like PCR. It can detect single nucleotide polymorphisms. The process involves denaturing the DNA, annealing probes to the target sequence, and ligating the probes if they match, repeating for exponential amplification. Products are detected through gel electrophoresis or non-radioactive methods. LCR has applications in infectious disease detection, genotyping and mutation analysis.
This document discusses various methods of genetic transfer, including natural genetic transfer between organisms as well as technological methods developed to manipulate genes. It describes how donor DNA can enter a recipient cell and recombine, producing genetically distinct offspring. Several gene transfer technologies are then outlined, including microinjection, biolistics, calcium phosphate precipitation, lipofection, and electroporation. The document explains the basic mechanisms and applications of each method while also noting their limitations for different purposes like gene therapy. In the conclusion, it emphasizes that gene transfer technologies now allow relatively easy and accurate introduction of genes into target cells to potentially cure diseases.
The DEAE-dextran method is a technique for transfection of nucleic acids into cultured mammalian cells. It involves mixing the nucleic acid with the cationic polymer DEAE-dextran to form complexes via electrostatic interactions. These complexes are added to cells and uptake is induced by osmotic shock, then the cells are washed and assayed for gene expression. The method is easy, low-cost, and can transfect a variety of cell types, but efficiency is lower than other techniques and high DEAE-dextran concentrations can be toxic.
Genetically modified or transgenic plants are plants that have been modified using genetic engineering techniques to introduce new traits. This document discusses the history and process of creating transgenic plants. It describes how transgenic plants are generated by transferring genes from other species into the target plant using either indirect methods like Agrobacterium-mediated transformation or direct physical methods like biolistics. The document provides details on the various applications of transgenic plants including producing herbicide resistance, insect resistance, virus resistance, and improving nutritional quality.
- Lambda bacteriophage cloning vectors were developed to overcome limitations in the size of DNA that could be inserted into unmodified lambda vectors.
- Segments of the non-essential lambda genome could be deleted to allow insertion of up to 18kb of new DNA while still allowing packaging.
- Natural selection was used to generate lambda strains lacking restriction sites, allowing restriction-based cloning.
- The first lambda vectors were insertion and replacement vectors, while later cosmids allowed cloning of fragments up to 52kb.
This document describes various methods for transferring genes into organisms. Biological methods include using viruses like cauliflower mosaic virus (CaMV) to transfer genes into plants. Physical methods include electroporation, which uses electric pulses to create pores in cell membranes through which DNA can enter. Liposomes and direct methods like microinjection and particle bombardment can also be used to directly transfer DNA. Chemical methods involve using compounds like polyethylene glycol (PEG) to destabilize cell membranes and allow DNA uptake. While physical methods can target single cells, they may damage cells. Biological methods using vectors are often more efficient but less controlled. Overall the document provides an overview of the key gene transfer techniques.
introduction
What is virus
What is virus resistance plant
History
Gene use for develop virus resistance plant
Coat protein gene
cDNA of satellite RNA
Defective viral genome
Antisense RNA approach and
Ribozyme – mediated protection
conclusion
References
The document summarizes the ligase chain reaction (LCR) technique. LCR amplifies DNA sequences using four probes and ligase and polymerase enzymes instead of amplifying DNA through nucleotide polymerization like PCR. It can detect single nucleotide polymorphisms. The process involves denaturing the DNA, annealing probes to the target sequence, and ligating the probes if they match, repeating for exponential amplification. Products are detected through gel electrophoresis or non-radioactive methods. LCR has applications in infectious disease detection, genotyping and mutation analysis.
This document discusses various methods of genetic transfer, including natural genetic transfer between organisms as well as technological methods developed to manipulate genes. It describes how donor DNA can enter a recipient cell and recombine, producing genetically distinct offspring. Several gene transfer technologies are then outlined, including microinjection, biolistics, calcium phosphate precipitation, lipofection, and electroporation. The document explains the basic mechanisms and applications of each method while also noting their limitations for different purposes like gene therapy. In the conclusion, it emphasizes that gene transfer technologies now allow relatively easy and accurate introduction of genes into target cells to potentially cure diseases.
The DEAE-dextran method is a technique for transfection of nucleic acids into cultured mammalian cells. It involves mixing the nucleic acid with the cationic polymer DEAE-dextran to form complexes via electrostatic interactions. These complexes are added to cells and uptake is induced by osmotic shock, then the cells are washed and assayed for gene expression. The method is easy, low-cost, and can transfect a variety of cell types, but efficiency is lower than other techniques and high DEAE-dextran concentrations can be toxic.
Genetically modified or transgenic plants are plants that have been modified using genetic engineering techniques to introduce new traits. This document discusses the history and process of creating transgenic plants. It describes how transgenic plants are generated by transferring genes from other species into the target plant using either indirect methods like Agrobacterium-mediated transformation or direct physical methods like biolistics. The document provides details on the various applications of transgenic plants including producing herbicide resistance, insect resistance, virus resistance, and improving nutritional quality.
- Lambda bacteriophage cloning vectors were developed to overcome limitations in the size of DNA that could be inserted into unmodified lambda vectors.
- Segments of the non-essential lambda genome could be deleted to allow insertion of up to 18kb of new DNA while still allowing packaging.
- Natural selection was used to generate lambda strains lacking restriction sites, allowing restriction-based cloning.
- The first lambda vectors were insertion and replacement vectors, while later cosmids allowed cloning of fragments up to 52kb.
Gene transfer technology pharmacology biotechnology basic methods
Natural, physical, chemical methods of gene transfer.
Along with advantages and limitations, and applications.
This document provides information about zinc finger proteins. It begins with an introduction to zinc finger motifs, which are protein structural domains characterized by the coordination of zinc ions. The document then discusses the history of zinc finger discovery, functions, and families. It provides details on the most common Cys2His2 zinc finger proteins and their role in DNA recognition and transcriptional regulation. The document also examines uses of zinc finger nucleases for genome editing and their mechanism of action involving creating double-strand breaks in DNA.
The document discusses the production of transgenic organisms. It defines key terms like transgenic, transgene, and transgenesis. It explains that a transgene is a foreign gene deliberately inserted into an organism's genome, making it transgenic. The common methods to produce transgenic animals are pronuclear microinjection and embryonic stem cell methods. The document provides examples of important transgenic animals and their applications in medicine, agriculture, and research.
This document discusses the use of Agrobacterium tumefaciens for genetic transformation of plants through its Ti plasmid, which contains tumor-inducing (Ti) DNA that can integrate genes for traits like herbicide and insect resistance into plant genomes. The process involves A. tumefaciens attaching to wounded plant cells and inducing virulence genes that cause the Ti plasmid and its T-DNA to integrate into the plant's nuclear DNA, resulting in transformed cells that can be used to generate transgenic crops with desirable traits.
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
Agrobacterium mediated Transformation-Mechanism of gene transfer,Virulence induction in presence of Plant secondary metabolites,Chromosomal genes and Vir genes,Agrobacterium tumefaciens – pathogen and useful tool for genetic engineering
This document discusses various types of vectors used for cloning and expressing plant genes, including their characteristics and uses. It describes cloning vectors, expression vectors, and integration vectors. Common vector types that are discussed in detail include plasmids, bacterial viruses (bacteriophages), cosmids, and plant viruses. The binary vector system and co-integration vector strategy for transforming plants using Agrobacterium tumefaciens are also summarized.
Direct Gene Transfer method (gene gun method).ShaistaKhan60
Direct gene transfer methods rely on delivering naked DNA directly into plant cells without the use of Agrobacterium. Biolistic or gene gun particle bombardment is a direct gene transfer method where gold or tungsten particles are coated with DNA and shot into plant tissue using a gene gun. The DNA can then integrate into the plant genome. The method involves creating recombinant plasmids with the gene of interest, coating them onto microcarriers, bombarding embryogenic plant callus, selecting transformed cells, and regenerating plants. Transgenic papaya developed using the gene gun method were resistant to papaya ring spot virus.
This document describes the transformation of competent E. coli cells using the CaCl2 method. It defines competent cells as cells that can uptake exogenous DNA and discusses plasmids, which are self-replicating DNA that can carry useful genes. The document explains that transformation can be natural or artificial, and the artificial process involves making cells competent through ice-cold CaCl2 treatment, then applying heat shock to induce the cells to uptake exogenous DNA like plasmids.
The document discusses various gene transfer techniques, including direct and indirect methods. Direct methods involve physical techniques like electroporation, particle bombardment, and microinjection which directly transfer DNA into cells. Chemical methods also directly transfer DNA using liposomes, DEAE dextran, or calcium phosphate precipitation. Indirect methods involve vector-mediated transfer, including Agrobacterium-mediated transfer where bacteria insert DNA into plant cells, and virus-mediated transfer using viruses like tobacco mosaic virus or cauliflower mosaic virus.
Gene cloning involves copying a gene and inserting it into a self-replicating vector to produce multiple copies of the gene. PCR (polymerase chain reaction) is a technique used to amplify specific genes. Both gene cloning and PCR are important for obtaining pure samples of genes and amplifying them for various applications like sequencing, expression of proteins, and genetic engineering.
Genetic engineering,recombinant DNA technology..ganeshbond
1) In 1996, scientists in Edinburgh announced the creation of Dolly the sheep, the first mammal cloned from an adult cell.
2) Dolly's birth sparked debate around the controversial technique of cloning and its potential application to humans.
3) In 2001, scientists in Texas cloned the world's first kitten, named Cc, using a cell from an adult tortoiseshell cat. The kitten was unveiled in 2002.
Recombinant DNA technology involves combining DNA from different species and inserting it into a host organism. Major breakthroughs in the 1960s and 1970s included the discovery of restriction enzymes and reverse transcriptase, which allowed scientists to generate recombinant DNA molecules. This led to important applications in medicine, agriculture, and industry, such as the development of insulin produced through recombinant DNA in 1979. The technology has continued advancing, with achievements like the cloning of Dolly the sheep in 1997 and approval of drugs produced through recombinant DNA like Cinqair for treating asthma.
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.
Retroviral vectors are derived from wild type retroviruses like Moloney murine leukemia virus. They are engineered to carry foreign genes into target cells. The vectors contain cis-acting elements from the viral genome like LTRs and packaging signals but lack the trans-acting genes gag, pol, and env. This prevents replication but allows the vector and its gene of interest to integrate stably into the host cell genome. Retroviral vectors show promise for gene therapy applications but also have limitations like requiring actively dividing cells for transduction and potential risks of insertional mutagenesis leading to cancer.
Direct methods of gene transfer in rDNA technologySijo A
The area of electrofusion and electroporation of plant cells is rapidly expanding. Both techniques present the plant scientist with powerful tools for gene transfer. Further modifications and improvements of these technologies should extend their gene transfer capability to a broad range of plants.
Metabolic engineering involves redirecting enzymatic reactions in an organism to produce new compounds or improve existing ones. It focuses on intermediates or products like starch, vitamins, amino acids. Successful approaches introduce new pathways, like producing provitamin A in rice. Rate-limiting steps and multi-level modifications are important. Unexpected results can occur. Commercialization requires safety characterization. Goals include overproducing desired compounds, underproducing unwanted ones, and novel compounds. Engineering targets pathways for carbohydrates, amino acids, lipids, alkaloids, terpenoids and more. Important examples include high-lysine plants, nutritionally-improved cottonseed oil, and Golden Rice which produces beta-carotene in rice
Simian virus 40 (SV40) is a DNA virus that can cause tumors in monkeys and humans, and it was first identified as a contaminant in polio vaccines in the 1960s. SV40 has been widely used as a cloning vector due to its ability to efficiently deliver genes into a variety of cells without killing the host cell or eliciting an immune response. Future research prospects for SV40 vectors include developing recombinant versions for gene transfer applications and furthering understanding of related retroviruses.
The document discusses applications of recombinant DNA technology, focusing on important recombinant proteins and their uses. It provides details on the production of human insulin, interferons, and hepatitis B vaccine through recombinant DNA techniques. Human insulin was the first therapeutic protein produced via recombinant DNA, and is made by inserting the human insulin gene into E. coli bacteria. Interferons are produced recombinantly in yeast cells, which can properly glycosylate the proteins. The hepatitis B vaccine is made from antigenic proteins of the hepatitis B virus produced recombinantly, potentially through genetic engineering of banana plants.
The following powerpoint presentation consist of all the essential knowledge about recombinant DNA technology, its techniues and different uses, advantages, disadvantages,
This document provides an overview of bacterial genetics. It discusses how DNA carries genetic information that is transcribed into RNA and translated into proteins. It describes DNA and RNA structure, DNA replication, and extrachromosomal genetic elements like plasmids. It also summarizes various genetic mechanisms that allow transmission of genetic material between bacteria like transformation, transduction, conjugation, and transposition. The use of these genetic techniques in applications like genetic engineering, restriction endonucleases, DNA probes, blotting techniques, PCR, and molecular epidemiology is also covered at a high level.
Gene transfer technology pharmacology biotechnology basic methods
Natural, physical, chemical methods of gene transfer.
Along with advantages and limitations, and applications.
This document provides information about zinc finger proteins. It begins with an introduction to zinc finger motifs, which are protein structural domains characterized by the coordination of zinc ions. The document then discusses the history of zinc finger discovery, functions, and families. It provides details on the most common Cys2His2 zinc finger proteins and their role in DNA recognition and transcriptional regulation. The document also examines uses of zinc finger nucleases for genome editing and their mechanism of action involving creating double-strand breaks in DNA.
The document discusses the production of transgenic organisms. It defines key terms like transgenic, transgene, and transgenesis. It explains that a transgene is a foreign gene deliberately inserted into an organism's genome, making it transgenic. The common methods to produce transgenic animals are pronuclear microinjection and embryonic stem cell methods. The document provides examples of important transgenic animals and their applications in medicine, agriculture, and research.
This document discusses the use of Agrobacterium tumefaciens for genetic transformation of plants through its Ti plasmid, which contains tumor-inducing (Ti) DNA that can integrate genes for traits like herbicide and insect resistance into plant genomes. The process involves A. tumefaciens attaching to wounded plant cells and inducing virulence genes that cause the Ti plasmid and its T-DNA to integrate into the plant's nuclear DNA, resulting in transformed cells that can be used to generate transgenic crops with desirable traits.
MBB 501 PLANT BIOTECHNOLOGY
INFORMATION ABOUT DIFFERENT DNA MODIFYING ENZYMES
WHAT IS AN ENZYME?
Alkaline Phosphatase
Polynucleotide kinase
Terminal deoxyneucleotidyl transferase
Nucleases
Exonuclease
Bal31 Exonuclease III
Endonuclease
S1 endonulease
Deoxyribonuclease 1 (Dnase 1)
RNase A
RNase H
Restriction Endonuclease
PvuI
PvuII
Different types of endonuclease enzymes
The recognition sequences for some of the most frequently used restriction endonucleases.
Categorization of enzymes
Isoschizomers
Neoschizomers
Isocaudomers
Agrobacterium mediated Transformation-Mechanism of gene transfer,Virulence induction in presence of Plant secondary metabolites,Chromosomal genes and Vir genes,Agrobacterium tumefaciens – pathogen and useful tool for genetic engineering
This document discusses various types of vectors used for cloning and expressing plant genes, including their characteristics and uses. It describes cloning vectors, expression vectors, and integration vectors. Common vector types that are discussed in detail include plasmids, bacterial viruses (bacteriophages), cosmids, and plant viruses. The binary vector system and co-integration vector strategy for transforming plants using Agrobacterium tumefaciens are also summarized.
Direct Gene Transfer method (gene gun method).ShaistaKhan60
Direct gene transfer methods rely on delivering naked DNA directly into plant cells without the use of Agrobacterium. Biolistic or gene gun particle bombardment is a direct gene transfer method where gold or tungsten particles are coated with DNA and shot into plant tissue using a gene gun. The DNA can then integrate into the plant genome. The method involves creating recombinant plasmids with the gene of interest, coating them onto microcarriers, bombarding embryogenic plant callus, selecting transformed cells, and regenerating plants. Transgenic papaya developed using the gene gun method were resistant to papaya ring spot virus.
This document describes the transformation of competent E. coli cells using the CaCl2 method. It defines competent cells as cells that can uptake exogenous DNA and discusses plasmids, which are self-replicating DNA that can carry useful genes. The document explains that transformation can be natural or artificial, and the artificial process involves making cells competent through ice-cold CaCl2 treatment, then applying heat shock to induce the cells to uptake exogenous DNA like plasmids.
The document discusses various gene transfer techniques, including direct and indirect methods. Direct methods involve physical techniques like electroporation, particle bombardment, and microinjection which directly transfer DNA into cells. Chemical methods also directly transfer DNA using liposomes, DEAE dextran, or calcium phosphate precipitation. Indirect methods involve vector-mediated transfer, including Agrobacterium-mediated transfer where bacteria insert DNA into plant cells, and virus-mediated transfer using viruses like tobacco mosaic virus or cauliflower mosaic virus.
Gene cloning involves copying a gene and inserting it into a self-replicating vector to produce multiple copies of the gene. PCR (polymerase chain reaction) is a technique used to amplify specific genes. Both gene cloning and PCR are important for obtaining pure samples of genes and amplifying them for various applications like sequencing, expression of proteins, and genetic engineering.
Genetic engineering,recombinant DNA technology..ganeshbond
1) In 1996, scientists in Edinburgh announced the creation of Dolly the sheep, the first mammal cloned from an adult cell.
2) Dolly's birth sparked debate around the controversial technique of cloning and its potential application to humans.
3) In 2001, scientists in Texas cloned the world's first kitten, named Cc, using a cell from an adult tortoiseshell cat. The kitten was unveiled in 2002.
Recombinant DNA technology involves combining DNA from different species and inserting it into a host organism. Major breakthroughs in the 1960s and 1970s included the discovery of restriction enzymes and reverse transcriptase, which allowed scientists to generate recombinant DNA molecules. This led to important applications in medicine, agriculture, and industry, such as the development of insulin produced through recombinant DNA in 1979. The technology has continued advancing, with achievements like the cloning of Dolly the sheep in 1997 and approval of drugs produced through recombinant DNA like Cinqair for treating asthma.
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.
Retroviral vectors are derived from wild type retroviruses like Moloney murine leukemia virus. They are engineered to carry foreign genes into target cells. The vectors contain cis-acting elements from the viral genome like LTRs and packaging signals but lack the trans-acting genes gag, pol, and env. This prevents replication but allows the vector and its gene of interest to integrate stably into the host cell genome. Retroviral vectors show promise for gene therapy applications but also have limitations like requiring actively dividing cells for transduction and potential risks of insertional mutagenesis leading to cancer.
Direct methods of gene transfer in rDNA technologySijo A
The area of electrofusion and electroporation of plant cells is rapidly expanding. Both techniques present the plant scientist with powerful tools for gene transfer. Further modifications and improvements of these technologies should extend their gene transfer capability to a broad range of plants.
Metabolic engineering involves redirecting enzymatic reactions in an organism to produce new compounds or improve existing ones. It focuses on intermediates or products like starch, vitamins, amino acids. Successful approaches introduce new pathways, like producing provitamin A in rice. Rate-limiting steps and multi-level modifications are important. Unexpected results can occur. Commercialization requires safety characterization. Goals include overproducing desired compounds, underproducing unwanted ones, and novel compounds. Engineering targets pathways for carbohydrates, amino acids, lipids, alkaloids, terpenoids and more. Important examples include high-lysine plants, nutritionally-improved cottonseed oil, and Golden Rice which produces beta-carotene in rice
Simian virus 40 (SV40) is a DNA virus that can cause tumors in monkeys and humans, and it was first identified as a contaminant in polio vaccines in the 1960s. SV40 has been widely used as a cloning vector due to its ability to efficiently deliver genes into a variety of cells without killing the host cell or eliciting an immune response. Future research prospects for SV40 vectors include developing recombinant versions for gene transfer applications and furthering understanding of related retroviruses.
The document discusses applications of recombinant DNA technology, focusing on important recombinant proteins and their uses. It provides details on the production of human insulin, interferons, and hepatitis B vaccine through recombinant DNA techniques. Human insulin was the first therapeutic protein produced via recombinant DNA, and is made by inserting the human insulin gene into E. coli bacteria. Interferons are produced recombinantly in yeast cells, which can properly glycosylate the proteins. The hepatitis B vaccine is made from antigenic proteins of the hepatitis B virus produced recombinantly, potentially through genetic engineering of banana plants.
The following powerpoint presentation consist of all the essential knowledge about recombinant DNA technology, its techniues and different uses, advantages, disadvantages,
This document provides an overview of bacterial genetics. It discusses how DNA carries genetic information that is transcribed into RNA and translated into proteins. It describes DNA and RNA structure, DNA replication, and extrachromosomal genetic elements like plasmids. It also summarizes various genetic mechanisms that allow transmission of genetic material between bacteria like transformation, transduction, conjugation, and transposition. The use of these genetic techniques in applications like genetic engineering, restriction endonucleases, DNA probes, blotting techniques, PCR, and molecular epidemiology is also covered at a high level.
Recombinant DNA technology involves manipulating DNA from different sources to produce novel DNA molecules. It has several key steps: isolating the desired DNA and vector, joining them using enzymes to create recombinant DNA, introducing this into a host cell, and selecting cells that express the gene. This technology has many applications including producing human insulin and growth hormones through bacteria, developing vaccines by cloning genes for antigens, and creating monoclonal antibodies. It allows mass production of important biological substances that were previously difficult to obtain.
Biotechnology can be summarized as follows:
1. It uses living organisms or their components to develop useful products. This includes genetically modifying microbes, plants and animals.
2. Key branches include genetic engineering, tissue culture, DNA fingerprinting and gene therapy. Genetic engineering is used to create GMOs by transferring genes between organisms.
3. Important tools for genetic engineering include vectors, restriction enzymes, DNA ligases, and host cells used to replicate recombinant DNA. This allows genes of interest to be isolated and transferred to create GMOs.
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.
DNA cloning is the process of making multiple, identical copies of a particular piece of DNA. In a typical DNA cloning procedure, the gene or other DNA fragment of interest (perhaps a gene for a medically important human protein) is first inserted into a circular piece of DNA called a plasmid.- [https://www.khanacademy.org/science/...dna.../dna-cloning.../a/overview-dna-cloning]
Genetic transformation & success of DNA ligation Sabahat Ali
DNA is ligated through DNA Ligase, problems may occur during DNA ligation are
1) vector cyclization
2) vector-vector concatemers
3) target DNA-target DNA ligation
Genetic engineering involves manipulating genetic material (DNA) to achieve desired goals in predetermined ways. Recombinant DNA technology joins DNA from different species, which are inserted into a host organism to produce new genetic combinations valuable to science, medicine, agriculture and industry. This process involves generating DNA fragments, selecting desired fragments, inserting them into cloning vectors, introducing the vectors into host cells, multiplying the cells containing recombinant DNA, and expressing genes to produce desired products.
This document discusses various gene transfer techniques used in genetic engineering. It describes direct techniques like chemically stimulated DNA uptake using polyethylene glycol (PEG), transduction using bacteriophages, electroporation using high voltage electricity, and microinjection of DNA into fertilized eggs. It also discusses indirect techniques like microprojectile bombardment which shoots DNA-coated particles into plant cells, and Agrobacterium-mediated transfer where the bacterium transfers tumor-inducing (T-DNA) from its Ti plasmid into the host plant genome.
Biotechnology refers to the use of living organisms or their components to develop products or perform processes for specific use. Recombinant DNA technology uses genetic engineering techniques to create recombinant DNA by cutting and joining DNA molecules containing different genetic material. This allows genes to be transferred between organisms for applications such as producing human insulin or diagnosing diseases. The key steps involve isolating DNA, using restriction enzymes to cut the DNA at specific sites, joining DNA fragments using DNA ligase, and inserting the recombinant DNA into a host cell where it can be replicated through bacterial transformation.
This document provides an overview of genetic engineering and its applications to microorganisms. It defines genetic engineering as the direct manipulation of an organism's genome using biotechnology. The key steps involved are isolating the gene of interest, inserting it into a vector, introducing the vector into a host cell, and harvesting the gene product from the clone. Common hosts used are bacteria, yeast, plant and animal cells. The document also discusses some tools used in genetic engineering like restriction enzymes and DNA ligase. It outlines several applications of genetic engineering in medicine, research, agriculture and industry. It concludes by noting some ethical and safety concerns regarding genetically modified organisms.
This document provides an overview of recombinant DNA technology. It begins by describing the central dogma of molecular biology - DNA is transcribed into RNA which is translated into protein. It then discusses various applications of recombinant DNA technology, including gene isolation, sequencing, PCR, gene therapy, and genetically modified crops. The document goes on to describe common techniques used in recombinant DNA, including restriction enzymes, vectors, transformation of host cells, and plasmid cloning. It provides examples of commonly used plasmid and phage vectors.
Cloning involves producing genetically identical copies of biological material such as DNA, cells, or whole organisms. The main steps of DNA cloning are: 1) cutting the DNA fragment to be cloned and the vector DNA with the same restriction enzymes, 2) joining the fragment to the vector via ligation, 3) introducing the recombinant DNA into host cells via transformation, 4) selecting transformed cells using antibiotic resistance markers on the vector, and 5) screening clones to identify those containing the inserted DNA fragment. Common cloning vectors include plasmids, which are small extra-chromosomal DNA molecules that replicate within host bacteria or yeast cells.
Recombinant DNA technology allows DNA from different species to be isolated, cut, and spliced together to form new recombinant molecules. Key tools for recombinant DNA technology include restriction enzymes, ligases, polymerases, vectors, and host cells. Recombinant DNA technology has many applications, including producing human insulin and other proteins for medical use, genetically engineering plants for crop improvement, and DNA fingerprinting for criminal investigations.
This document provides an overview of biotechnology, including definitions, principles, processes, tools, and applications. It defines biotechnology as using biological agents like microorganisms and enzymes to provide goods and services for human welfare. The principles of biotechnology draw from fields like microbiology, genetics, biochemistry, and chemical engineering. Key processes discussed include genetic engineering, recombinant DNA technology using tools like restriction enzymes, cloning vectors, the polymerase chain reaction, and competent host organisms like E. coli. Applications of biotechnology highlighted are in health care through recombinant proteins, industrial biotechnology, plant biotechnology, and environmental biotechnology.
Genetic engineering and Transformation methodsManjunath R
Genetic engineering involves manipulating an organism's genome using modern DNA technology. This document discusses various genetic transformation methods, including both direct and indirect methods. Indirect methods involve using Agrobacterium tumefaciens to transfer DNA into plant cells. Direct methods discussed include particle bombardment, polyethylene glycol treatment, electroporation, and microinjection. The document provides details on the process, mechanisms, applications and history of genetic engineering and transformation techniques.
This document provides an overview of DNA cloning. It discusses taking a gene of interest from a source DNA, inserting it into a vector such as a plasmid, and using this recombinant DNA to transform bacteria. This allows the gene of interest to be replicated in large quantities. Key steps include using restriction enzymes to cut the DNA pieces for ligation, transforming bacteria with the recombinant plasmid, and selecting for bacteria containing the cloned gene insert. The goal of cloning is to generate multiple copies of a gene for study and protein production.
Recombinant DNA technology allows for the isolation, cloning, and manipulation of genes. Two key advances enabled this field: genetic engineering using restriction enzymes to isolate and modify genes in vitro, and DNA sequencing to determine the order of nucleotides. Recombinant DNA is generated by joining DNA from different sources, and molecular cloning produces large quantities of a particular DNA fragment through construction of a recombinant vector, introduction into a host cell, selective propagation of cells containing the vector, and extraction of the cloned DNA.
Transformation is the process of altering an organism's genetic makeup by inserting new genes. Common transformation methods include Agrobacterium-mediated transformation, particle bombardment, protoplast transformation using polyethylene glycol or electroporation, and fibre-mediated DNA delivery. Agrobacterium transformation involves the bacteria transferring T-DNA from its Ti plasmid into the plant genome, while direct methods introduce naked DNA into plant cells using physical methods like particle bombardment or chemical treatments that make cell membranes permeable. Transformation allows improving crop traits like yield and stress resistance.
This document provides an overview of DNA cloning. It begins by defining cloning as making identical copies of DNA, genes, or cells. The basic steps of DNA cloning are described, including using a source DNA, vector, restriction enzymes to cut DNA, ligation to join DNA fragments, transformation of host bacteria, and selection of recombinant clones. Common vectors like plasmids are discussed along with selection techniques like blue-white screening. The document emphasizes that the goal is to generate multiple copies of the cloned insert DNA. Examples are given of important medical and agricultural applications of cloning genes.
Here is the updated list of Top Best Ayurvedic medicine for Gas and Indigestion and those are Gas-O-Go Syp for Dyspepsia | Lavizyme Syrup for Acidity | Yumzyme Hepatoprotective Capsules etc
Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
8 Surprising Reasons To Meditate 40 Minutes A Day That Can Change Your Life.pptxHolistified Wellness
We’re talking about Vedic Meditation, a form of meditation that has been around for at least 5,000 years. Back then, the people who lived in the Indus Valley, now known as India and Pakistan, practised meditation as a fundamental part of daily life. This knowledge that has given us yoga and Ayurveda, was known as Veda, hence the name Vedic. And though there are some written records, the practice has been passed down verbally from generation to generation.
Adhd Medication Shortage Uk - trinexpharmacy.comreignlana06
The UK is currently facing a Adhd Medication Shortage Uk, which has left many patients and their families grappling with uncertainty and frustration. ADHD, or Attention Deficit Hyperactivity Disorder, is a chronic condition that requires consistent medication to manage effectively. This shortage has highlighted the critical role these medications play in the daily lives of those affected by ADHD. Contact : +1 (747) 209 – 3649 E-mail : sales@trinexpharmacy.com
These lecture slides, by Dr Sidra Arshad, offer a quick overview of the physiological basis of a normal electrocardiogram.
Learning objectives:
1. Define an electrocardiogram (ECG) and electrocardiography
2. Describe how dipoles generated by the heart produce the waveforms of the ECG
3. Describe the components of a normal electrocardiogram of a typical bipolar lead (limb II)
4. Differentiate between intervals and segments
5. Enlist some common indications for obtaining an ECG
6. Describe the flow of current around the heart during the cardiac cycle
7. Discuss the placement and polarity of the leads of electrocardiograph
8. Describe the normal electrocardiograms recorded from the limb leads and explain the physiological basis of the different records that are obtained
9. Define mean electrical vector (axis) of the heart and give the normal range
10. Define the mean QRS vector
11. Describe the axes of leads (hexagonal reference system)
12. Comprehend the vectorial analysis of the normal ECG
13. Determine the mean electrical axis of the ventricular QRS and appreciate the mean axis deviation
14. Explain the concepts of current of injury, J point, and their significance
Study Resources:
1. Chapter 11, Guyton and Hall Textbook of Medical Physiology, 14th edition
2. Chapter 9, Human Physiology - From Cells to Systems, Lauralee Sherwood, 9th edition
3. Chapter 29, Ganong’s Review of Medical Physiology, 26th edition
4. Electrocardiogram, StatPearls - https://www.ncbi.nlm.nih.gov/books/NBK549803/
5. ECG in Medical Practice by ABM Abdullah, 4th edition
6. Chapter 3, Cardiology Explained, https://www.ncbi.nlm.nih.gov/books/NBK2214/
7. ECG Basics, http://www.nataliescasebook.com/tag/e-c-g-basics
2. Biotechnology
• The use of microorganisms, cells, or cell components to make
a product
– Foods that are produced by the action of microorganisms
(bacteria and yeasts)
– Antibiotics
– Vitamins
– Enzymes
3. 1. Mutation & Selection
– Genetic mutation and recombination provide a diversity of
organisms.
– The process of natural selection allows the growth of those
best adapted to a given environment.
2. Microorganisms can exchange genes in a process of natural DNA
recombination – genetic modification.
What do we know?
4. • Modern Biotechnology
– Selection
• Culture a naturally-occurring microbe that produces desired product (antibiotic producing
bacteria)
– Mutation and selection
• Mutagens cause mutations that might result in a microbe with a desirable trait (penicillin
produced by the fungus over 1000 times)
• Select and culture microbe with the desired mutation
– Gene modification
• Change a specific DNA bases ( change the corresponding codons) to change a protein
• Recombinant DNA Technology:
– Recombinant DNA - DNA that has been artificially manipulated to combine
genes from two different sources.
– Genes transferred - among unrelated species via laboratory manipulation.
– Genetic engineering - human manipulation of an organism's genetic material in
a way that does not occur under natural conditions
What could we do?
5. Figure 9.1.2
An Overview of Recombinant DNA Technologies
1. Gene of interest (DNA) is isolated
(DNA fragment)
2. A desired gene is inserted into a
DNA molecule - vector
(plasmid, bacteriophage or a viral genome)
3. The vector inserts the DNA into a
new cell, which is grown to form a
clone.
(bacteria, yeast, plant or animal cell)
4. Large quantities of the gene product
can be harvested from the clone.
6. Tools for Genetic engineering
1. Restriction Enzymes
• Naturally produced by bacteria – restriction endonucleases
– Natural function - destroy bacteriophage DNA in bacterial cells
– Cannot digest host DNA with methylated C (cytosine)
• A restriction enzyme
– Substrate –DNA -recognizes one particular nucleotide sequence in DNA
and cuts the DNA molecule (breaks down the bond between two
nucleotides)
• Prepackaged kits are available for rDNA techniques
sticky ends blunt ends
8. Figure 9.2
Restriction Enzymes
• Fragments of DNA produced by the same restriction enzyme will
spontaneously join by base pairing.
• Each of the DNA strands will have a break
9. Tools for Genetic engineering
2. Ligase
• DNA ligase is a enzyme that can link together DNA strands that
have double-strand breaks (a break in both complementary strands
of DNA).
– Naturally DNA ligase has applications in both DNA replication and DNA
repair .
– Needs ATP
• DNA ligase has extensive use in molecular biology laboratories
for genetic recombination experiments
ATP
10. Tools for Genetic engineering
3. Vectors
• Vectors - small pieces of DNA used for cloning (the gene to be
inserted into the genetically modified organism must be combined
with other genetic elements in order for it to work properly)
• Requirements of the Vector
1. Self-replication - able to replicate in the host (origin of
repliction)
2. Cloning site (site for recognition of restriction nucleases)
3. Promoter (and operator) - to support the gene (new DNA) expression in the
host
4. Selectable marker – antibiotic resistance
5. Proper size
11. Vectors
1. Plasmid vectors
– Plasmids are self-replicating circular molecules of DNA
– Encode antibiotic resistance ( selection marker)
2. Viral vectors - retroviruses, adenoviruses and herpes viruses
– Accept much larger pieces of DNA
– Mammalian hosts
12. 1. Bacteria
- E. coli - used because is easily grown and its
genomics are well understood.
– Gene product is purified from host cells
2. Yeasts - Saccharomyces cerevisiae
– Used because it is easily grown and its genomics are known
– May express eukaryotic genes easily
– Continuously secrete the gene product.
– Easily collected and purified
3. Plant cells and whole plants
– May express eukaryotic genes easily
– Plants are easily grown - produce plants with new properties.
4. Mammalian cells
– May express eukaryotic genes easily
– Harder to grow
– Medical use.
Hosts for DNA recombinant technology
13. • The recombinant vector carrying foreign
DNA needs to be transferred into the
suitable host cells. Several methods have
been developed for introduction of
recombinant DNA molecule into host cells.
• According to types of vectors and host
cells, the methods are adopted. Some of
the methods of gene transfer into host
cells are briefly discussed below:
14. Insert the naked DNA into a host cell
1.Transformation
treatment make cells competent to accept foreign DNA (CaCl2 make
pores in cell membrane)
2. Electroporation
use electrical current to form microscopic pores in the membranes of
cell
3. Microinjection
4. Gene gun
Figure 9.5b
15. 1. Transformation :
• Introduction of rDNA molecules into a
living cell is called transformation. The
DNA molecule comes in the contact of cell
surface. Then it is taken up by the host
cells.
• However, in nature the frequency of
transformation of many cells (e.g. yeast
and mammalian cells) is very less.
Secondly, all the time host cells do not
undergo transformation. Because they are
not prepared for it.
16.
17. • A temperature sock of either low
temperature (0-5°C) or high temperature
(37- 45°C) is required for transformation.
Mandel and Higa (1970) reported that E.
coli cells become competent to uptake
DNA by treating cells with ice cold
CaCl2 and exposing the cells to 42°C for
about 90 seconds.
18. 2. Transduction :
• The action or process
of transducing; especially : the transfer of
genetic material from one microorganism
to another by a viral agent (such as a
bacteriophage)
19.
20. 3. Electro oration (Electric
Field-mediated Membrane
Permeation) :
• In electroporation an electric current at high
voltage (about 350 V) is applied in a solution
containing foreign DNA and fragile host cells.
This creates transient microscopic pores in cell
membrane of naked protoplasts.
• Consequently foreign DNA enters into the
protoplast through these pores. The transformed
protoplasts are cultured in vitro which
regenerate respective cell walls.
21.
22. 4. Microinjection :
• In this technique foreign DNA is directly and forcibly
injected into the nucleus of animal and plant cells
through a glass micropipette containing very fine tip of
about 0.5 mm diameter.
• It resembles with injection needle. In 1982, for the first
time Rubin and Spradling introduced Drosophila gene
into P-element and microinjected into embryo. In 1982,
R.D. Palmiter and R.L.
• Brinter introduced rabbit recombinant pBR322 containing
a growth hormone gene into mature embryo of mouse in
vitro. The microinjected embryos were transferred into
uterus of mice which gave birth to transgenic mice.
23.
24. 5. Particle Bombardment Gun
(Biolistics) :
• This technique was developed by Stanford
in 1987. In this method macroscopic gold
or tungusten particles are coated with
desired DNA. A plastic micro-carrier
containing DNA coated gold/tungusten
particles are placed near rupture disc.
• The particles are bombarded onto target
cells by the bombardment apparatus.
Consequently foreign DNA is forcibly
delivered into the host cells.
25.
26. CALCIUM PHOSPHATE MEDIATED DNA TRANSFER
• The process of transfection involves the admixture of isolated DNA
(10-100ug) with solution of calcium chloride and potassium
phosphate under condition which allow the precipitate of calcium
phosphate to be formed.
• Cells are then incubated with precipitated DNA either in solution or
in tissue culture dish. A fraction of cells will take up the calcium
phosphate DNA precipitate by endocytosis.
• Transfection efficiencies using calcium phosphate can be quite low,
in the range of 1-2 %. It can be increased if very high purity DNA is
used and the precipitate allowed to form slowly.
27.
28.
29. Limitations of calcium phosphate mediated DNA transfer
•Frequency is very low.
•Integrated genes undergo substantial modification.
•Many cells do not like having the solid precipitate adhering
to them and the surface of their culture vessel.
•Due to above limitations transfection applied to somatic gene
therapy is limited.
30.
31.
32. Recombinant DNA technology - Cloning
A process of producing genetically modified organisms
A multi-step process.
1. Isolating and copying the genetic material of interest (DNA
fragment ).
2. Building a construct (recombinant DNA - vector and desired gene)
containing all the genetic elements for correct expression.
3. Inserting the vector into the host organism, directly through
injection or transformation.
4. Selecting the cells expressing that gene by growing under positive
selection (of an antibiotic or chemical) – clone .
5. Growing successfully the clone (transformed organisms).
33. Blue-white screening system
2. The vector is then transformed into host competent cell
(bacteria).
• Host is sensitive to ampicillin
• Host is β-galactosidase negative (do not carry LacZ gene)
3. The transformed cells are grown in the presence of:
• ampicillin.
• X-gal – substrate for β-galactosidase
– a colourless modified galactose sugar
– When metabolized by β-galactosidase form an insoluble
product (5-bromo-4 chloroindole) which is bright blue, and
thus functions as an indicator
4. Results
– Clones lacking the vector will not grow.
– Clones containing the vector without the new gene will be resistant to
ampicillin, able to metabolized X-gal and will be blue.
– Clones containing the recombinant vector will be resistant to
ampicillin and unable to hydrolyze X-gal (white colonies).
• If the ligation was successful, the bacterial colony will be
white; if not, the colony will be blue.
34. Obtaining DNA – gene of interest
1.Gene libraries are made of pieces of
an entire genome stored in plasmids
or phages
2. cDNA (complementary DNA) is made
from mRNA by reverse transcriptase
(enzyme found in retroviruses)
- Intron free DNA
3. Synthetic DNA is made by a DNA synthesis machine
35. Screening for the desired Gene
• Identify the particular cell that contains the specific gene of interest
(Presence of the vector with correct gene of interest)
• A short piece of labeled
DNA called a DNA probe
can be used to identify clones
carrying the desired gene.
• Radioactive labeled
• Fluorescent labeled
Labeled DNA probe ( 32P or fluorescence)
5’ *AGGCTTGTACTTTGGCGG 3’
36. Copying the genetic material of interest - PCR
• Polymerase Chain Reaction (PCR)
– A reaction to make multiple copies of a piece of DNA enzymatically
• Polymerase – enzyme is DNA polymerase from Thermus aquaticus – Taq
polymerase
– Taq's optimum temperature for activity is 75-80°C
– Can replicate a 1000 base pair strand of DNA in less than 10 seconds at
72°C
• Chain – chain of cycles of multiplication
DNA-
Dissociation temperature
Hybridization temperature
37. PCR amplify DNA to detectable levels
Figure 9.4.1
PCR reaction mixture :
1. Target DNA (template)
2. Short primers- to hybridize to the 5’ end
of each DNA strand
3. four NTP – ATP, GTP, TTP, and CTP
4. Buffer
5. DNA Taq Polymerase (enzyme)
Cycle program in PCR machine:
1. Denaturation -95ºC
2. Annealing (hybridization)- 60-65 ºC
3. Polymerase reaction -72 ºC
Product
Start with 1 molecule
First cycle – 2 molecules
Second cycle – 4 molecules
…
….
Finished after 30 cycles – 1,073,741,842 molecules
30 cycles
38. DNA sequencing
• DNA sequencing - is the process of
determining the precise order of nucleotides
within a DNA molecule
( A, G, C and T in a molecule of DNA)
39. Applications of recombinant DNA technology
1. Scientific applications
– Many copies of DNA can be produced
– Increase understanding of DNA
– Identify mutations in DNA
– Alter the phenotype of an organism
– Bioinformatics is the use of computer applications to study
genetic data;
– Proteomics – proteomics is the study of a cell’s proteins.
• determination of all the proteins expressing in the cell
40. Applications of recombinant DNA technology
• Shotgun sequencing - Recombinant DNA techniques were used
to map the human genome through the Human Genome Project
- has 24 distinct chromosomes (22 autosomal + X + Y)
- with a total of approximately 3 billion DNA base pairs
– containing an estimated 20,000–25,000 genes
– with only about 1.5-2% coding for proteins
– the rest comprised by RNA genes, regulatory
sequences, introns and controversially so-called
junk DNA
– This provides tools for diagnosis and
possibly the repair of genetic diseases
41. Applications of recombinant DNA technology
2. Diagnose genetic disease
– RFLP analysis (Restriction fragment
length polymorphism)
• DNA profiling involved restriction
enzyme digestion, followed by
Southern blot analysis
– Southern blotting is used for
detection of a specific DNA sequence
in DNA sample
• DNA probes can be used to quickly
identify a pathogen in body tissue or
food.
– PCR analysis with specific primers
43. 3. Recombinant DNA techniques can be used to for genetic
fingerprinting identification
• Forensic microbiology - use
DNA fingerprinting to identify
the source of bacterial or viral
pathogens.
– bioterrorism attacks (Anthrax in U.S. Mail)
– medical negligence (Tracing HIV to
a physician who injected it)
– outbreaks of foodborne diseases
Figure 9.16
Applications of recombinant DNA technology
44. 4. Agricultural Applications
– Cells from plants with desirable characteristics can be cloned to
produce many identical cells, then can be used to produce whole plants
from which seeds can be harvested.
– Some bacteria can transfer genes to unrelated species
• Agrobacterium tumefaciens - a plant pathogen
• Cause tumors in plants
• Natural genetic engineer
Applications of recombinant DNA technology
45. Genetic engineering manipulation
GMO
Site of insertion foreign DNA
Selection
• Genes for resistance to herbicide glyphosate, Bt toxin, and
pectinase suppression have been engineered into crop plants.
• Genetically modified Rhizobium has enhanced nitrogen
fixation.
• Genetically modified Pseudomonas is a biological insecticide
that produces Bacillus thuringiensis toxin.
46. 5. Nanotechnology
• Bacteria can make molecule-sized particles
– Bacillus cells growing on selenium form chains of elemental selenium
Applications of recombinant DNA technology
47. 6. Therapeutic Applications
– Produce human proteins – hormones and enzymes
• Insulin
• hGH
• INFα, INFβ and INFγ
– Vaccines
• Cells and viruses can be modified to produce a pathogen’s surface protein
– Influenza
– Hepatitis B
– Cervical cancer vaccine
• Nonpathogenic viruses carrying genes for pathogen’s antigens as DNA
vaccines
• DNA vaccines consist of circular rDNA
– Gene therapy can be used to cure genetic diseases by replacing
the defective or missing gene.
– Gene silencing – RNA interference - siRNA or microRNA
Applications of recombinant DNA technology
48. • Accidental release - strict safety standards are used to avoid it
– Some microbes used in recombinant DNA cloning have been altered so
that they cannot survive outside the laboratory.
– Microorganisms intended for use in the environment may be modified to
contain suicide genes so that the organisms do not persist in the
environment.
• Ethical questions
– Should employers and insurance companies have access to a person’s
genetic records?
– Will some people be targeted for either breeding or sterilization?
– Will genetic counseling be available to everyone?
• GMO - Genetically modified crops must be safe for consumption
and for release in the environment.
Safety Issues and Ethics
49. • Compare and contrast biotechnology, genetic modification, and recombinant DNA.
• Compare selection and mutation.
• Identify the roles of a clone and a vector in making recombined DNA.
• Define restriction enzymes, and outline how they are used to make recombinant DNA.
• Outline the steps in PCR and provide an example of its use.
• Describe how a gene library is made
• Differentiate cDNA from synthetic DNA.
• List the properties of vectors.
• Describe the use of plasmid and viral vectors.
• Describe five ways of getting DNA into a cell.
• Explain how each of the following are used to locate a clone: antibiotic-resistance
genes, DNA probes.
• List one advantage of engineering the following: E. coli, Saccharomyces cerevisiae,
mammalian cells, plant cells.
• Discuss the value of the Human Genome Project.
• Define the following terms: random shotgun sequencing, bioinformatics, proteomics.
• Diagram the Southern blot procedure and provide an example of its use.
• Diagram DNA fingerprinting and provide an example of its use.
• List the advantages of, and problems associated with, the use of genetic modification
techniques.
Learning objectives