This document discusses various gene transfer methods. It defines gene transfer as the insertion of genetic material into a cell. There are natural methods like conjugation, transformation, and transduction that involve the transfer of genes between bacteria. There are also artificial physical, chemical, and electrical methods to transfer genes into cells, including microinjection, gene guns, calcium phosphate, liposomes, and electroporation. The document provides examples of how these various gene transfer methods can be used to insert genes into bacteria, plants, and animals.
This document provides an overview of Agrobacterium-mediated gene transformation. It begins with an introduction to genetic transformation methods, including direct and indirect techniques. It then discusses Agrobacterium, including its classification, the history of using it for gene transformation, and features of its T-DNA and virulence genes. The document outlines the process of T-DNA transfer from Agrobacterium to plant cells. Finally, it describes some common methods for Agrobacterium-mediated gene transfer, such as infection through wounds, leaf disk, and co-cultivation techniques.
Electroporation is a method to transform cells by creating transient pores in the cell membrane through applying brief high-voltage electric pulses, allowing DNA to enter the cell. It involves suspending cells in a solution with DNA between electrodes and applying pulses of 4000-8000 V/cm for milliseconds. This forms pores in the membrane through which DNA can enter. It is commonly used to transform bacteria, yeast, plant protoplasts, and transfect eukaryotic cells. Key factors influencing electroporation include field strength, pulse length, DNA purity and concentration, and cell growth conditions.
This document summarizes Agrobacterium-mediated plant transformation. It describes how the soil bacterium Agrobacterium tumefaciens causes crown gall disease in plants by transferring oncogenic T-DNA from its Ti plasmid into the plant genome. Scientists have exploited this natural process to develop transformation systems where they insert new genes between the border sequences of disarmed Ti plasmids, allowing transfer of the recombinant T-DNA into plant cells. While effective in dicots, transformation of monocots proved more difficult due to their limited regeneration ability, though biolistic methods using microprojectile bombardment have succeeded in some important crop species.
Methods of Gene Transfer document discusses various methods of transferring genes into plants to create transgenic plants. It describes two main categories of gene transfer methods - physical and biological. Physical methods include microinjection, biolistics (gene gun), electroporation, and particle bombardment. Biological methods include Agrobacterium-mediated transformation, which involves using the bacteria Agrobacterium tumefaciens to transfer DNA into plant cells. The document also discusses transformation cassettes, selection of transgenic plants, analysis of transgenic plants, and some examples of commercially important transgenic crops like golden rice and Roundup Ready corn.
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.
Adenoviral vectors are modified adenoviruses that can deliver genetic material into host cells. Adenoviruses are medium-sized, non-enveloped viruses containing double-stranded DNA. They can efficiently transfer DNA/RNA into cells and have been used to construct viral vectors. Wild type adenoviruses are modified by deleting non-essential genes and adding exogenous genetic material to create viral vectors. Three generations of adenoviral vectors have been developed for gene therapy, with later generations having improved safety profiles and ability to carry larger DNA payloads.
This document provides an overview of Agrobacterium-mediated gene transformation. It begins with an introduction to genetic transformation methods, including direct and indirect techniques. It then discusses Agrobacterium, including its classification, the history of using it for gene transformation, and features of its T-DNA and virulence genes. The document outlines the process of T-DNA transfer from Agrobacterium to plant cells. Finally, it describes some common methods for Agrobacterium-mediated gene transfer, such as infection through wounds, leaf disk, and co-cultivation techniques.
Electroporation is a method to transform cells by creating transient pores in the cell membrane through applying brief high-voltage electric pulses, allowing DNA to enter the cell. It involves suspending cells in a solution with DNA between electrodes and applying pulses of 4000-8000 V/cm for milliseconds. This forms pores in the membrane through which DNA can enter. It is commonly used to transform bacteria, yeast, plant protoplasts, and transfect eukaryotic cells. Key factors influencing electroporation include field strength, pulse length, DNA purity and concentration, and cell growth conditions.
This document summarizes Agrobacterium-mediated plant transformation. It describes how the soil bacterium Agrobacterium tumefaciens causes crown gall disease in plants by transferring oncogenic T-DNA from its Ti plasmid into the plant genome. Scientists have exploited this natural process to develop transformation systems where they insert new genes between the border sequences of disarmed Ti plasmids, allowing transfer of the recombinant T-DNA into plant cells. While effective in dicots, transformation of monocots proved more difficult due to their limited regeneration ability, though biolistic methods using microprojectile bombardment have succeeded in some important crop species.
Methods of Gene Transfer document discusses various methods of transferring genes into plants to create transgenic plants. It describes two main categories of gene transfer methods - physical and biological. Physical methods include microinjection, biolistics (gene gun), electroporation, and particle bombardment. Biological methods include Agrobacterium-mediated transformation, which involves using the bacteria Agrobacterium tumefaciens to transfer DNA into plant cells. The document also discusses transformation cassettes, selection of transgenic plants, analysis of transgenic plants, and some examples of commercially important transgenic crops like golden rice and Roundup Ready corn.
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.
Adenoviral vectors are modified adenoviruses that can deliver genetic material into host cells. Adenoviruses are medium-sized, non-enveloped viruses containing double-stranded DNA. They can efficiently transfer DNA/RNA into cells and have been used to construct viral vectors. Wild type adenoviruses are modified by deleting non-essential genes and adding exogenous genetic material to create viral vectors. Three generations of adenoviral vectors have been developed for gene therapy, with later generations having improved safety profiles and ability to carry larger DNA payloads.
This document provides information on various methods of gene transfer in plants, including Agrobacterium-mediated gene transfer and direct gene transfer methods. Direct methods rely on delivering large amounts of DNA to plant cells through techniques like particle bombardment, electroporation, and microinjection. Agrobacterium-mediated gene transfer utilizes the bacterium Agrobacterium, which transfers genes into plant genomes. The document discusses several direct and Agrobacterium-mediated methods in detail and provides advantages and limitations of each approach.
This document discusses methods for producing haploid plants. It begins by defining haploid plants and their significance. It then describes the two main approaches for producing haploids - in vivo and in vitro. For in vivo, it outlines several techniques including androgenesis, gynogenesis, distant hybridization, and chemical/radiation treatments. For in vitro, it focuses on anther culture and microspore culture, providing details on the protocol for anther culture in tobacco including pre-treatment, culture conditions, and factors that influence success rates.
Agrobacterium is a soil-borne bacteria that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into plant cells. This process allows for efficient gene transfer. The T-DNA contains oncogenes that cause tumor formation in plants as well as genes for opine synthesis. Virulence genes on the Ti plasmid regulate T-DNA transfer through a type IV secretion system. There are two main strategies for using Agrobacterium for plant genetic engineering - co-integration vectors, where the gene of interest is inserted into the Ti plasmid, and binary vectors, where the T-DNA and virulence genes are on separate replicons that complement each other in Agrobacterium.
The document summarizes a seminar on the Ti plasmid. It discusses that the Ti plasmid is found in Agrobacterium tumefaciens and is responsible for crown gall tumor formation in plants. It describes the organization and structure of the Ti plasmid, including the T-DNA region flanked by borders that is transferred to plant cells. Two common vector systems used for plant transformation, the cointegrate vector and binary vector, are explained. The cointegrate vector involves integration of an intermediate vector with the Ti plasmid, while the binary vector separates the plasmid and virulence genes. Finally, the general process of Agrobacterium-mediated plant transformation is outlined.
Gene transfer techniques can be direct or indirect. Direct techniques introduce foreign DNA into plant cells without a biological agent, using methods like microinjection, microprojectiles, protoplast fusion, electroporation, and polyethylene glycol treatment. Indirect gene transfer uses the bacterium Agrobacterium tumefaciens, which transfers DNA (T-DNA) from its tumor-inducing plasmid into the host plant genome, allowing genetic modification of plants. Techniques like bacterial transformation and transduction can also directly transfer genes between bacteria using viruses or naked DNA. Overall, a variety of methods have been developed to introduce foreign genes into organisms and achieve genetic modification.
Gene transfer technologies can be used to treat diseases by inserting therapeutic genes into cells. There are viral and non-viral methods of gene transfer. Viral methods use viruses like retroviruses, adenoviruses, and adeno-associated viruses to efficiently deliver genes. Non-viral methods include mechanical techniques like electroporation, microinjection, and biolistics (gene gun), as well as chemical methods like liposomes, calcium phosphate, and polyethylene glycol. Each method has advantages and limitations for different applications in research and potential gene therapy.
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Androgenesis is the production of haploid plants through the culture of male gametophytes or microspores. There are two main methods - anther culture and isolated pollen/microspore culture. Anther culture involves excising anthers from flower buds and culturing them on nutrient media, while microspore culture isolates microspores from anthers. Several factors influence androgenesis success, including genotype, anther wall components, culture medium, growth regulators, and physical conditions. Androgenic haploids can develop directly from microspores or indirectly through a callus phase, following various developmental pathways. Androgenesis allows for the efficient production of haploid plants for breeding programs.
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
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.
Agrobacterium tumefaciens is commonly used to genetically modify dicot plants through its ability to transfer DNA to plant cells. It causes crown gall disease by transferring oncogenic T-DNA from its tumor-inducing plasmid into wounded plant cells. The T-DNA encodes genes that cause tumor formation and the production of opines, which the bacteria can use as nutrients. The binary vector system was developed to overcome challenges with manipulating large Ti plasmids, allowing foreign genes to be stably introduced between the T-DNA borders and transferred to plant cells. Agrobacterium-mediated transformation is now widely used to produce transgenic plants.
Agrobacterium mediated gene transfer in plants.ICHHA PURAK
This power point presentation consist of 41 slides. Attempts have been made to illustrate how Agrobacterium behaves us natural genetic engineer. How it can infect a plant through wound and a part of DNA present on Ti plasmid is Tranferred and causes disease as crown gall in the infected plant. In second part of the presentation attempts have been made to describe how Agrobacterium can be utilized for iinsertion of desired gene into the plant,what manipulation are to be made with Agrobacterium.How infection and transfer of desired gene can be made possible.What is the role of plant tissue culture etc.
application of plant transformation for productivity and performanceAshika Raveendran
This document discusses pathogenesis-related (PR) proteins in plants. It notes that plants produce PR proteins when under pathogen attack, like from fungi. There are over 14 families of PR proteins with different functions, such as having antifungal, glucanase, or chitinase activity. The document outlines several specific PR proteins (PR-1 through PR-5), describing their molecular weights, presence in various plant species, and antifungal mechanisms. It also mentions several other types of antifungal proteins found in plants.
The document discusses organogenesis, which is the development of adventitious organs or primordial from undifferentiated plant cell mass through differentiation. It describes the process, including dedifferentiation and redifferentiation stages. There are two types of organogenesis - direct organogenesis which does not involve callus formation, and indirect organogenesis which involves callus formation. Organogenesis is used in plant tissue culture to regenerate plants through shoot or root cultures and is influenced by factors like explant source and size, plant growth regulators, and culture conditions. It has commercial applications in micropropagation of plants.
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It does this by transferring a segment of its tumor-inducing plasmid (Ti plasmid) called T-DNA into the plant's DNA. The T-DNA contains genes that cause uncontrolled cell growth. Researchers developed techniques using the Ti plasmid and Agrobacterium to genetically transform plants by replacing the tumor-causing genes with desired genes. This involves either a binary vector system with the T-DNA on a separate small plasmid, or co-integration of a new plasmid containing the gene of interest into the Ti plasmid. Transformed plants can be regenerated from infected plant cells or tissues.
The document discusses the production of double haploid plants through anther and pollen culture techniques. It provides background on the history of double haploid development, the importance of double haploids in plant breeding, and methods used to induce haploids including anther culture, pollen culture, ovary slice culture, and ovule culture. Key factors influencing anther culture success are also reviewed, such as genotype, culture medium, microspore stage, temperature, and donor plant physiology. Advantages and disadvantages of generating double haploid lines are presented.
The document discusses selectable marker genes that are commonly used in plant transformation systems. Selectable marker genes are included in transformation vectors along with the target gene of interest. They confer resistance to transformed cells when grown on media containing toxic substances like antibiotics, herbicides, or antimetabolites. This allows transformed cells to survive while non-transformed cells die. There are three main categories of selectable marker genes: antibiotic resistance genes, antimetabolite marker genes, and herbicide resistance genes. Common examples of genes used include nptII for kanamycin resistance, pat/bar for phosphinothricin/glufosinate resistance, and epsps/aroA for glyphosate resistance.
This document discusses various gene transfer methods. It defines gene transfer as the insertion of genetic material into a cell. There are natural methods like conjugation, transformation, and transduction that involve the transfer of genes between bacteria. There are also artificial physical, chemical, and electrical methods to transfer genes into bacteria, plants, and animals. These include microinjection, biolistics, calcium phosphate, liposomes, and electroporation. The document provides details on how each of these methods work and their advantages and limitations.
Gene transfer techniques can be used to transfer genes between organisms. There are natural methods like conjugation, transformation, and transduction that transfer genes between bacteria. Artificial methods like microinjection, biolistics, calcium phosphate transfection, liposome transfection, and electroporation can be used to transfer genes into both bacteria and eukaryotic cells. Agrobacterium mediated transfer is used to transfer genes into plant cells and involves the T-DNA region of the Ti plasmid. The transferred gene is then integrated into the host genome.
This document provides information on various methods of gene transfer in plants, including Agrobacterium-mediated gene transfer and direct gene transfer methods. Direct methods rely on delivering large amounts of DNA to plant cells through techniques like particle bombardment, electroporation, and microinjection. Agrobacterium-mediated gene transfer utilizes the bacterium Agrobacterium, which transfers genes into plant genomes. The document discusses several direct and Agrobacterium-mediated methods in detail and provides advantages and limitations of each approach.
This document discusses methods for producing haploid plants. It begins by defining haploid plants and their significance. It then describes the two main approaches for producing haploids - in vivo and in vitro. For in vivo, it outlines several techniques including androgenesis, gynogenesis, distant hybridization, and chemical/radiation treatments. For in vitro, it focuses on anther culture and microspore culture, providing details on the protocol for anther culture in tobacco including pre-treatment, culture conditions, and factors that influence success rates.
Agrobacterium is a soil-borne bacteria that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into plant cells. This process allows for efficient gene transfer. The T-DNA contains oncogenes that cause tumor formation in plants as well as genes for opine synthesis. Virulence genes on the Ti plasmid regulate T-DNA transfer through a type IV secretion system. There are two main strategies for using Agrobacterium for plant genetic engineering - co-integration vectors, where the gene of interest is inserted into the Ti plasmid, and binary vectors, where the T-DNA and virulence genes are on separate replicons that complement each other in Agrobacterium.
The document summarizes a seminar on the Ti plasmid. It discusses that the Ti plasmid is found in Agrobacterium tumefaciens and is responsible for crown gall tumor formation in plants. It describes the organization and structure of the Ti plasmid, including the T-DNA region flanked by borders that is transferred to plant cells. Two common vector systems used for plant transformation, the cointegrate vector and binary vector, are explained. The cointegrate vector involves integration of an intermediate vector with the Ti plasmid, while the binary vector separates the plasmid and virulence genes. Finally, the general process of Agrobacterium-mediated plant transformation is outlined.
Gene transfer techniques can be direct or indirect. Direct techniques introduce foreign DNA into plant cells without a biological agent, using methods like microinjection, microprojectiles, protoplast fusion, electroporation, and polyethylene glycol treatment. Indirect gene transfer uses the bacterium Agrobacterium tumefaciens, which transfers DNA (T-DNA) from its tumor-inducing plasmid into the host plant genome, allowing genetic modification of plants. Techniques like bacterial transformation and transduction can also directly transfer genes between bacteria using viruses or naked DNA. Overall, a variety of methods have been developed to introduce foreign genes into organisms and achieve genetic modification.
Gene transfer technologies can be used to treat diseases by inserting therapeutic genes into cells. There are viral and non-viral methods of gene transfer. Viral methods use viruses like retroviruses, adenoviruses, and adeno-associated viruses to efficiently deliver genes. Non-viral methods include mechanical techniques like electroporation, microinjection, and biolistics (gene gun), as well as chemical methods like liposomes, calcium phosphate, and polyethylene glycol. Each method has advantages and limitations for different applications in research and potential gene therapy.
Presented by- MD JAKIR HOSSAIN
Doctoral Research Scholar
Department of Agricultural Genetic Engineering ,
Faculty of Agricultural Sciences and Technologies,
Nigde Omer Halisdemir University, Turkey
E. Mail- mjakirbotru@gmail.com
Androgenesis is the production of haploid plants through the culture of male gametophytes or microspores. There are two main methods - anther culture and isolated pollen/microspore culture. Anther culture involves excising anthers from flower buds and culturing them on nutrient media, while microspore culture isolates microspores from anthers. Several factors influence androgenesis success, including genotype, anther wall components, culture medium, growth regulators, and physical conditions. Androgenic haploids can develop directly from microspores or indirectly through a callus phase, following various developmental pathways. Androgenesis allows for the efficient production of haploid plants for breeding programs.
What are an expression vector? Detailed description of plant gene structure. Plant expression vector systems are generally consists of Ri and Ti plasmids.
The other vectors which are generally used are DNA and RNA viruses.
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.
Agrobacterium tumefaciens is commonly used to genetically modify dicot plants through its ability to transfer DNA to plant cells. It causes crown gall disease by transferring oncogenic T-DNA from its tumor-inducing plasmid into wounded plant cells. The T-DNA encodes genes that cause tumor formation and the production of opines, which the bacteria can use as nutrients. The binary vector system was developed to overcome challenges with manipulating large Ti plasmids, allowing foreign genes to be stably introduced between the T-DNA borders and transferred to plant cells. Agrobacterium-mediated transformation is now widely used to produce transgenic plants.
Agrobacterium mediated gene transfer in plants.ICHHA PURAK
This power point presentation consist of 41 slides. Attempts have been made to illustrate how Agrobacterium behaves us natural genetic engineer. How it can infect a plant through wound and a part of DNA present on Ti plasmid is Tranferred and causes disease as crown gall in the infected plant. In second part of the presentation attempts have been made to describe how Agrobacterium can be utilized for iinsertion of desired gene into the plant,what manipulation are to be made with Agrobacterium.How infection and transfer of desired gene can be made possible.What is the role of plant tissue culture etc.
application of plant transformation for productivity and performanceAshika Raveendran
This document discusses pathogenesis-related (PR) proteins in plants. It notes that plants produce PR proteins when under pathogen attack, like from fungi. There are over 14 families of PR proteins with different functions, such as having antifungal, glucanase, or chitinase activity. The document outlines several specific PR proteins (PR-1 through PR-5), describing their molecular weights, presence in various plant species, and antifungal mechanisms. It also mentions several other types of antifungal proteins found in plants.
The document discusses organogenesis, which is the development of adventitious organs or primordial from undifferentiated plant cell mass through differentiation. It describes the process, including dedifferentiation and redifferentiation stages. There are two types of organogenesis - direct organogenesis which does not involve callus formation, and indirect organogenesis which involves callus formation. Organogenesis is used in plant tissue culture to regenerate plants through shoot or root cultures and is influenced by factors like explant source and size, plant growth regulators, and culture conditions. It has commercial applications in micropropagation of plants.
Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in plants. It does this by transferring a segment of its tumor-inducing plasmid (Ti plasmid) called T-DNA into the plant's DNA. The T-DNA contains genes that cause uncontrolled cell growth. Researchers developed techniques using the Ti plasmid and Agrobacterium to genetically transform plants by replacing the tumor-causing genes with desired genes. This involves either a binary vector system with the T-DNA on a separate small plasmid, or co-integration of a new plasmid containing the gene of interest into the Ti plasmid. Transformed plants can be regenerated from infected plant cells or tissues.
The document discusses the production of double haploid plants through anther and pollen culture techniques. It provides background on the history of double haploid development, the importance of double haploids in plant breeding, and methods used to induce haploids including anther culture, pollen culture, ovary slice culture, and ovule culture. Key factors influencing anther culture success are also reviewed, such as genotype, culture medium, microspore stage, temperature, and donor plant physiology. Advantages and disadvantages of generating double haploid lines are presented.
The document discusses selectable marker genes that are commonly used in plant transformation systems. Selectable marker genes are included in transformation vectors along with the target gene of interest. They confer resistance to transformed cells when grown on media containing toxic substances like antibiotics, herbicides, or antimetabolites. This allows transformed cells to survive while non-transformed cells die. There are three main categories of selectable marker genes: antibiotic resistance genes, antimetabolite marker genes, and herbicide resistance genes. Common examples of genes used include nptII for kanamycin resistance, pat/bar for phosphinothricin/glufosinate resistance, and epsps/aroA for glyphosate resistance.
This document discusses various gene transfer methods. It defines gene transfer as the insertion of genetic material into a cell. There are natural methods like conjugation, transformation, and transduction that involve the transfer of genes between bacteria. There are also artificial physical, chemical, and electrical methods to transfer genes into bacteria, plants, and animals. These include microinjection, biolistics, calcium phosphate, liposomes, and electroporation. The document provides details on how each of these methods work and their advantages and limitations.
Gene transfer techniques can be used to transfer genes between organisms. There are natural methods like conjugation, transformation, and transduction that transfer genes between bacteria. Artificial methods like microinjection, biolistics, calcium phosphate transfection, liposome transfection, and electroporation can be used to transfer genes into both bacteria and eukaryotic cells. Agrobacterium mediated transfer is used to transfer genes into plant cells and involves the T-DNA region of the Ti plasmid. The transferred gene is then integrated into the host genome.
This document discusses various techniques for gene transfer, including natural methods like conjugation, transformation, and transduction, as well artificial methods like microinjection, biolistics, calcium phosphate transfection, liposome-mediated transfection, and electroporation. It provides details on how each method works, such as how conjugation involves transfer of DNA between bacteria via sex pili, how transformation involves direct DNA uptake by competent bacteria, and how transduction involves transfer of DNA between bacteria via bacteriophages. The document also discusses Agrobacterium-mediated plant transformation and applications of gene transfer techniques.
This document discusses various techniques for gene transfer, including natural methods like conjugation, transformation, and transduction, as well artificial methods like microinjection, biolistics, calcium phosphate and liposome mediated transfer, and electroporation. It provides details on how each method works, such as how conjugation involves transfer of DNA between bacteria via sex pili, and how electroporation uses electrical pulses to create pores in cell membranes to allow DNA entry. The document also summarizes screening and applications of transgenic techniques.
This document discusses various techniques for transferring genes, including natural and artificial methods. Natural methods include conjugation, transformation, transduction, and Agrobacterium-mediated transfer. Artificial methods include microinjection, biolistics, calcium phosphate transfection, liposome-mediated transfer, and electroporation. The document provides detailed descriptions of conjugation, transformation, transduction, Agrobacterium-mediated transfer, microinjection, biolistics, calcium phosphate transfection, liposome-mediated transfer, and electroporation. It also discusses screening methods for transgenes and applications of gene transfer techniques.
1. There are two main methods of gene transfer - direct and indirect gene transfer. Indirect transfer uses Agrobacterium-mediated transformation while direct transfer uses physical or chemical methods.
2. Agrobacterium-mediated transformation uses Agrobacterium tumefaciens to transfer T-DNA containing the gene of interest into the plant genome. The process involves co-cultivation of plant explants with Agrobacterium followed by selection and regeneration of transgenic plants.
3. Direct physical methods include biolistic transformation, microinjection, electroporation, and macroinjection. Direct chemical methods include PEG-mediated, calcium phosphate co-precipitation, and liposome-mediated transformation
Gene transfer technology pharmacology biotechnology basic methods
Natural, physical, chemical methods of gene transfer.
Along with advantages and limitations, and applications.
Electroporation uses electric pulses to create temporary pores in the cell membrane, allowing DNA entry. DNA-coated microprojectiles are accelerated into cells using a gene gun. Microinjection precisely inserts DNA into cells through fine glass needles. Calcium phosphate precipitation forms DNA-calcium phosphate complexes taken up by cells. Cationic liposomes fuse with cell membranes, transferring DNA across. Adenoviruses and retroviruses can deliver DNA to dividing and non-dividing cells. Agrobacterium transfers tumor-inducing (T-DNA) from its Ti plasmid into plant cells at wound sites.
This document provides an overview of various gene transfer tools and techniques. It discusses vector-mediated methods like Agrobacterium and viral vectors as well as direct or vector-less methods such as electroporation, biolistics, microinjection, liposome mediated, and calcium phosphate mediated gene transfer. For each method, it describes the basic process and provides some key details and applications. It also notes some advantages and limitations of different techniques. The document aims to inform readers about the various options available for inserting genes into plant cells.
Hi, I am RAFi ,student of Genetic Engineering and Biotechnology , Jashore university of science & Technology. It is my first uploading slide in slideshare.I am so glad for doing this work.
Gene transfer methods in animals can be natural or artificial. Natural methods include conjugation, transformation, and transduction which transfer genes between bacteria. Artificial methods like microinjection, biolistics, liposome mediated transfer, calcium phosphate mediated transfer, and electroporation are used to directly insert genes into cells. These techniques transfer genes into organisms for genetic engineering applications such as producing transgenic animals, developing vaccines, and gene therapy to treat diseases.
This document discusses methods for plant genetic engineering and transformation. It describes three main methods: electroporation, biolistics/particle bombardment, and Agrobacterium-mediated transformation. Agrobacterium transformation uses Agrobacterium tumefaciens bacteria to insert foreign DNA into plant cells. It involves removing crown gall genes from the Ti plasmid and replacing them with genes of interest. The process then involves inserting the modified Ti plasmid into Agrobacterium, mixing it with plant cells, and regenerating genetically modified plantlets.
Gene therapy involves introducing genetic material into human cells to treat diseases. The document discusses various gene transfer techniques including physical methods like electroporation and microinjection, chemical methods using calcium phosphate or cationic lipids, and biological methods using viral vectors. It also covers the goals and ideal characteristics of gene therapy as well as applications and future perspectives, concluding that gene therapy is a promising strategy but delivery of genes to target cells remains a key challenge.
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.
Transfection involves introducing foreign DNA into host cells to produce a new phenotype. There are two main methods of transfection - vector-mediated and non-vector mediated. Vector-mediated transfection uses bacteriophage, retroviral, cosmid, baculovirus, and plasmid vectors to introduce DNA. Non-vector mediated methods include direct techniques like microinjection, electroporation, and particle bombardment, and indirect techniques like calcium phosphate precipitation and DEAE-dextran. Retroviral vectors are modified retroviruses that can introduce foreign DNA into host chromosomal DNA. Microinjection involves injecting DNA directly into cells using a micropipette under a microscope. Electroporation uses electric pulses to create temporary
This document discusses the production of transgenic animals and plants. It describes three main methods for producing transgenic animals: DNA microinjection, retrovirus-mediated gene transfer, and embryonic stem cell-mediated gene transfer. It also discusses 11 methods for transforming plants, including Agrobacterium-mediated transformation, biolistic transformation, and floral dip transformation. Finally, it lists some beneficial traits that have been engineered in transgenic plants, such as stress tolerance, herbicide tolerance, and increased nutritional quality.
This document discusses various vectorless and direct gene transfer methods for recombinant DNA technology. It describes 10 different methods: chemical methods, electroporation, particle bombardment, lipofection, microinjection, macroinjection, pollen transformation, delivery via growing pollen tubes, laser induced transformation, and fibre mediated transformation. Each method directly introduces DNA into host cells without the use of biological vectors. The document provides details on the mechanisms and procedures for several of these direct gene transfer techniques.
There are three main modes of gene transfer: transformation, transfection, and transduction. Transformation involves the natural uptake of foreign DNA by a cell. Transfection is the deliberate introduction of genetic material into animal cells. Transduction uses viruses to transfer genes between bacterial cells. It can occur through a lysogenic or lytic phase. Conjugation is also discussed, which involves the direct contact and temporary exchange of genetic material between two bacterial cells.
When I was asked to give a companion lecture in support of ‘The Philosophy of Science’ (https://shorturl.at/4pUXz) I decided not to walk through the detail of the many methodologies in order of use. Instead, I chose to employ a long standing, and ongoing, scientific development as an exemplar. And so, I chose the ever evolving story of Thermodynamics as a scientific investigation at its best.
Conducted over a period of >200 years, Thermodynamics R&D, and application, benefitted from the highest levels of professionalism, collaboration, and technical thoroughness. New layers of application, methodology, and practice were made possible by the progressive advance of technology. In turn, this has seen measurement and modelling accuracy continually improved at a micro and macro level.
Perhaps most importantly, Thermodynamics rapidly became a primary tool in the advance of applied science/engineering/technology, spanning micro-tech, to aerospace and cosmology. I can think of no better a story to illustrate the breadth of scientific methodologies and applications at their best.
Phenomics assisted breeding in crop improvementIshaGoswami9
As the population is increasing and will reach about 9 billion upto 2050. Also due to climate change, it is difficult to meet the food requirement of such a large population. Facing the challenges presented by resource shortages, climate
change, and increasing global population, crop yield and quality need to be improved in a sustainable way over the coming decades. Genetic improvement by breeding is the best way to increase crop productivity. With the rapid progression of functional
genomics, an increasing number of crop genomes have been sequenced and dozens of genes influencing key agronomic traits have been identified. However, current genome sequence information has not been adequately exploited for understanding
the complex characteristics of multiple gene, owing to a lack of crop phenotypic data. Efficient, automatic, and accurate technologies and platforms that can capture phenotypic data that can
be linked to genomics information for crop improvement at all growth stages have become as important as genotyping. Thus,
high-throughput phenotyping has become the major bottleneck restricting crop breeding. Plant phenomics has been defined as the high-throughput, accurate acquisition and analysis of multi-dimensional phenotypes
during crop growing stages at the organism level, including the cell, tissue, organ, individual plant, plot, and field levels. With the rapid development of novel sensors, imaging technology,
and analysis methods, numerous infrastructure platforms have been developed for phenotyping.
ANAMOLOUS SECONDARY GROWTH IN DICOT ROOTS.pptxRASHMI M G
Abnormal or anomalous secondary growth in plants. It defines secondary growth as an increase in plant girth due to vascular cambium or cork cambium. Anomalous secondary growth does not follow the normal pattern of a single vascular cambium producing xylem internally and phloem externally.
Unlocking the mysteries of reproduction: Exploring fecundity and gonadosomati...AbdullaAlAsif1
The pygmy halfbeak Dermogenys colletei, is known for its viviparous nature, this presents an intriguing case of relatively low fecundity, raising questions about potential compensatory reproductive strategies employed by this species. Our study delves into the examination of fecundity and the Gonadosomatic Index (GSI) in the Pygmy Halfbeak, D. colletei (Meisner, 2001), an intriguing viviparous fish indigenous to Sarawak, Borneo. We hypothesize that the Pygmy halfbeak, D. colletei, may exhibit unique reproductive adaptations to offset its low fecundity, thus enhancing its survival and fitness. To address this, we conducted a comprehensive study utilizing 28 mature female specimens of D. colletei, carefully measuring fecundity and GSI to shed light on the reproductive adaptations of this species. Our findings reveal that D. colletei indeed exhibits low fecundity, with a mean of 16.76 ± 2.01, and a mean GSI of 12.83 ± 1.27, providing crucial insights into the reproductive mechanisms at play in this species. These results underscore the existence of unique reproductive strategies in D. colletei, enabling its adaptation and persistence in Borneo's diverse aquatic ecosystems, and call for further ecological research to elucidate these mechanisms. This study lends to a better understanding of viviparous fish in Borneo and contributes to the broader field of aquatic ecology, enhancing our knowledge of species adaptations to unique ecological challenges.
What is greenhouse gasses and how many gasses are there to affect the Earth.moosaasad1975
What are greenhouse gasses how they affect the earth and its environment what is the future of the environment and earth how the weather and the climate effects.
Travis Hills' Endeavors in Minnesota: Fostering Environmental and Economic Pr...Travis Hills MN
Travis Hills of Minnesota developed a method to convert waste into high-value dry fertilizer, significantly enriching soil quality. By providing farmers with a valuable resource derived from waste, Travis Hills helps enhance farm profitability while promoting environmental stewardship. Travis Hills' sustainable practices lead to cost savings and increased revenue for farmers by improving resource efficiency and reducing waste.
The ability to recreate computational results with minimal effort and actionable metrics provides a solid foundation for scientific research and software development. When people can replicate an analysis at the touch of a button using open-source software, open data, and methods to assess and compare proposals, it significantly eases verification of results, engagement with a diverse range of contributors, and progress. However, we have yet to fully achieve this; there are still many sociotechnical frictions.
Inspired by David Donoho's vision, this talk aims to revisit the three crucial pillars of frictionless reproducibility (data sharing, code sharing, and competitive challenges) with the perspective of deep software variability.
Our observation is that multiple layers — hardware, operating systems, third-party libraries, software versions, input data, compile-time options, and parameters — are subject to variability that exacerbates frictions but is also essential for achieving robust, generalizable results and fostering innovation. I will first review the literature, providing evidence of how the complex variability interactions across these layers affect qualitative and quantitative software properties, thereby complicating the reproduction and replication of scientific studies in various fields.
I will then present some software engineering and AI techniques that can support the strategic exploration of variability spaces. These include the use of abstractions and models (e.g., feature models), sampling strategies (e.g., uniform, random), cost-effective measurements (e.g., incremental build of software configurations), and dimensionality reduction methods (e.g., transfer learning, feature selection, software debloating).
I will finally argue that deep variability is both the problem and solution of frictionless reproducibility, calling the software science community to develop new methods and tools to manage variability and foster reproducibility in software systems.
Exposé invité Journées Nationales du GDR GPL 2024
hematic appreciation test is a psychological assessment tool used to measure an individual's appreciation and understanding of specific themes or topics. This test helps to evaluate an individual's ability to connect different ideas and concepts within a given theme, as well as their overall comprehension and interpretation skills. The results of the test can provide valuable insights into an individual's cognitive abilities, creativity, and critical thinking skills
EWOCS-I: The catalog of X-ray sources in Westerlund 1 from the Extended Weste...Sérgio Sacani
Context. With a mass exceeding several 104 M⊙ and a rich and dense population of massive stars, supermassive young star clusters
represent the most massive star-forming environment that is dominated by the feedback from massive stars and gravitational interactions
among stars.
Aims. In this paper we present the Extended Westerlund 1 and 2 Open Clusters Survey (EWOCS) project, which aims to investigate
the influence of the starburst environment on the formation of stars and planets, and on the evolution of both low and high mass stars.
The primary targets of this project are Westerlund 1 and 2, the closest supermassive star clusters to the Sun.
Methods. The project is based primarily on recent observations conducted with the Chandra and JWST observatories. Specifically,
the Chandra survey of Westerlund 1 consists of 36 new ACIS-I observations, nearly co-pointed, for a total exposure time of 1 Msec.
Additionally, we included 8 archival Chandra/ACIS-S observations. This paper presents the resulting catalog of X-ray sources within
and around Westerlund 1. Sources were detected by combining various existing methods, and photon extraction and source validation
were carried out using the ACIS-Extract software.
Results. The EWOCS X-ray catalog comprises 5963 validated sources out of the 9420 initially provided to ACIS-Extract, reaching a
photon flux threshold of approximately 2 × 10−8 photons cm−2
s
−1
. The X-ray sources exhibit a highly concentrated spatial distribution,
with 1075 sources located within the central 1 arcmin. We have successfully detected X-ray emissions from 126 out of the 166 known
massive stars of the cluster, and we have collected over 71 000 photons from the magnetar CXO J164710.20-455217.
The use of Nauplii and metanauplii artemia in aquaculture (brine shrimp).pptxMAGOTI ERNEST
Although Artemia has been known to man for centuries, its use as a food for the culture of larval organisms apparently began only in the 1930s, when several investigators found that it made an excellent food for newly hatched fish larvae (Litvinenko et al., 2023). As aquaculture developed in the 1960s and ‘70s, the use of Artemia also became more widespread, due both to its convenience and to its nutritional value for larval organisms (Arenas-Pardo et al., 2024). The fact that Artemia dormant cysts can be stored for long periods in cans, and then used as an off-the-shelf food requiring only 24 h of incubation makes them the most convenient, least labor-intensive, live food available for aquaculture (Sorgeloos & Roubach, 2021). The nutritional value of Artemia, especially for marine organisms, is not constant, but varies both geographically and temporally. During the last decade, however, both the causes of Artemia nutritional variability and methods to improve poorquality Artemia have been identified (Loufi et al., 2024).
Brine shrimp (Artemia spp.) are used in marine aquaculture worldwide. Annually, more than 2,000 metric tons of dry cysts are used for cultivation of fish, crustacean, and shellfish larva. Brine shrimp are important to aquaculture because newly hatched brine shrimp nauplii (larvae) provide a food source for many fish fry (Mozanzadeh et al., 2021). Culture and harvesting of brine shrimp eggs represents another aspect of the aquaculture industry. Nauplii and metanauplii of Artemia, commonly known as brine shrimp, play a crucial role in aquaculture due to their nutritional value and suitability as live feed for many aquatic species, particularly in larval stages (Sorgeloos & Roubach, 2021).
2. Gene
• A gene is the basic unit of heredity in a living
organism
• Genes are working subunit of DNA/RNA
3. Gene Transfer
– The insertion of genetic material into a cell
– It is defined simply as a technique to efficiently
and stably introduce foreign genes into the genome
of target cells
– The insertion of unrelated, therapeutic genetic
information in the form of DNA into target cells
Bacteria (or) Virus
Animal
Plant
6. Applications of Gene Transfer- Plants
• Provide resistance against viruses.
• Acquire insecticidal resistance.
• To strengthen the plant to grow against bacterial diseases.
• Develop the plants to grow in draught.
• Engineering plants for nutritional quality.
• Make the plants to grow in various seasons.
• Herbicide resistant plant can be made.
• Resistance against fungal pathogens.
• Engineering of plants for abiotic stress tolerance.
• Delayed ripening can be done.
7. • Clinical gene transfer applications
• Vaccine Development
• Production of transgenic animals
• Treatment of Cancer, AIDS
• Gene Discovery
• Gene Therapy
• GMO
Applications of Gene Transfer- Animals
11. Conjugation
• Requires the presence of a special plasmid called
the F plasmid.
• Bacteria that have a F plasmid are referred to as
as F+ or male. Those that do not have an F
plasmid are F- of female.
• The F plasmid consists of 25 genes that mostly
code for production of sex pilli.
• A conjugation event occurs when the male cell
extends his sex pilli and one attaches to the
female.
12. Conjugation
• This attached pilus is a temporary cytoplasmic
bridge through which a replicating F plasmid
is transferred from the male to the female.
• When transfer is complete, the result is two
male cells.
• When the F+ plasmid is integrated within the
bacterial chromosome, the cell is called an Hfr
cell (high frequency of recombination cell).
14. Transformation
• Transformation is the direct uptake of exogenous
DNA from its surroundings and taken up through
the cell membrane .
• Transformation occurs naturally in some species
of bacteria, but it can also be effected by artificial
treatment in other species.
• Cells that have undergone this treatment are said
to be competent.
• Any DNA that is not integrated into he
chromosome will be degraded.
16. Transduction
• Gene transfer from a donor to a recipient by way
of a bacteriophage
• Two way of integration
• If the lysogenic cycle is adopted, the phage
chromosome is integrated (by covalent bonds)
into the bacterial chromosome, where it can
remain dormant for thousands of generation
• The lytic cycle leads to the production of new
phage particles which are released by lysis of the
host.
18. Vector Mediated Gene Transfer
• Carrying molecule related gene transfer
–Plant vector- CaMV, TMV
•Agrobacterium mediated gene
transfer
–Animal vector- Sv40, Pox Viral vector
–Bacterial vector- pSC 101
19. Agrobacterium Mediated Gene Transfer
• The Agrobacterium system was historically the first
successful plant transformation system, marking the
break through in plant Genetic engineering in 1983.
• Large plasmids (140–235 kbp) in these bacteria are
called tumour inducing (Ti plasmid) and root inducing
(Ri plasmid) respectively.
• Virulence genes aid in the transfer of T-DNA into the
host plant cell. Ti plasmid contains 35 vir genes
arranged in 8 operons.
• Integrated in the plant nuclear DNA at random site
20. Agrobacterium Mediated Gene Transfer
Advantages
• Agrobacterium is capable of infecting infect plant cells,
tissue and organs.
• Capable of transfer of large fragments of DNA very
efficiently
• Integration of T DNA is a relative precise process.
• The stability of gene transferred in excellent.
Limitations :
• Host specificity
• Somaclonal variation
• Slow regeneration
• Inability to transfer multiple genes
21.
22. SV 40
• Simian virus 40 vector belongs to family polyoma
virus family.
• This virus induces tumours to animal cells and used
for gene transfer into eukaryotic cells
• Sv40 diameter is 45nm and its capsid contains 72
proteins
• It doesn’t have enzymes in structure
• Genome of sv40 is single molecule of double
stranded DNA
• It contain 5243bp in genome structure
24. Transposons
• Transposon, class of genetic elements that can
“jump” to different locations within a genome.
• In addition, most transposons eventually
become inactive and no longer move.
25. Microinjection
•DNA introduced into cells or protoplasts with use of very fine needles or
glass micropipettes
•T. P. Lin in 1966 (mouse eggs)
•The DNA solution is injected directly inside the cell using capillary glass
micropipettes with the help of micromanipulators of a microinjection
assembly.
•It is easier to use protoplast than cells since cell wall interferes with the
process of microinjection.
•The protoplast are usually immobilized in agarose (or) on a glass slides
coated with polylysine or by holding them under suction by a micropipette.
•The process of microinjection is technically demanding and time
consuming a maximum of 40-50 protoplasts can be microinjected in one
hour.
•The transformation frequency ranging between 14 to 66 %.
27. Microinjection
•Advantages
– Stable integration better than other method.
–DNA injected in this Process less modification only
occur
–Effective transforming primary cells and protoplast.
•Limitations
–Cost effective
–Random integration
–Only one cell at a time can be manipulated.
–Method not useful for the walled cells
–Skilled person required
28. Macroinjection
•DNA Macroinjection employing needles each with
diameter greater than cell diameter
•De la Pena et al., 1987
•The injection of DNA into the cells or tissues using
hypodermic syringe is known as macro injection.
•It is hypothesized that the DNA is taken up by
microspores during some specific stage of their
development.
•This approach is also very simple and easy; the only
problem concerns the frequency (0.07%) and the
consistency of stable transformants obtained.
31. Gene gun
• Biolistics method/Microprojectile/Particle bombardment
• Sanford and coworkers in 1987 (onion epidermal)
• Klein et al., transferred genomic RNA (TMV)
• The process of transformation employees foreign DNA
coated with (0.2-0.7 μm ) gold (or) are tungsten particles
to deliver into target plant cells.
• Two procedures have been used to this techniques
– i. By using pressurized helium gas.
– ii. By electro static energy released by a droplet of
water exposed to a high voltage.
33. Gene gun
• Advantages
– Need of protoplast obtaining can be avoided
– Walled intact cells can be penetrated
– Genome of subcellular organelles can be
manipulated.
• Limitations
– Random injection
– Need of equipment
– Damage cell
34. Laser
• It is method of introducing DNA into plant cells with a
laser micro beam.
• Small pores in the membrane are created by laser micro
beam.
• Lasers puncture holes in the cell membrane through
which DNA may enter into the cell cytoplasm.
• This method also used in Plant cells as well as Animal
cells.
• Lasers punctures holes in cell membrane through
which DNA may enter into the cell cytoplasm.
• But, There is no information/report on gene expression
or stable integration.
36. Sonication
• Sonoporation involves the use of ultrasound for temporary
permeabilization of the cell membrane allowing the uptake of
DNA, drugs or other therapeutic compounds from the
extracellular environment.
• This method leaves the compound trapped inside the cell after
ultrasound exposure.
• It employs of micro bubbles for enhancing the delivery of
large molecules like DNA. The micro bubbles form complex
with DNA followed by injection and ultrasound treatment to
deliver DNA into the target cells.
• Unlike other methods of transfection, sonoporation combines
the capability to enhance gene and drug transfer.
38. Sonication
Advantages
• Simple and highly efficient gene transfer method.
• No significant damage is cause to the target tissue.
Limitations
• Not suitable for tissues with open or complex
structures.
• High exposure to low-frequency (<MHz) ultrasounds
result in complete cellular death (rupture of the cell).
• Thus cellular viability must be taken into
consideration while employing this technique.
39. Calcium Phosphate
• Transfection is most common method
• Graham and Vander E.B. in 1973.
• 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 % .
• This technique is used for introducing DNA
into mammalian cells.
41. Advantages
–Mostly in mammalian cells.
–Along with other method
Limitations
–Small fraction of cells is stably integrated
–Frequency very Low
–Gene modification
–1-5% DNA enter into the nucleus.
–Toxic especially to primary cells
Calcium Phosphate
42. • This method is utilized for protoplast only.
• Polyethylene glycol stimulates endocytosis and
therefore DNA uptake occurs.
• Protoplasts are kept in the solution containing
polyethylene glycol (PEG).
• The molecular weight of PEG used is 8000
Dalton having the final concentration of 15%.
• CaCl2 or sucrose or Glucose – osmotic buffer
reagent .
• Mostly plant system (petunia)
PEG
43. • Advantages
–More efficient than electroporation
–Useful for plant
• Limitations
–Transformation not high
–Applicable only protoplast
–Increases nucleases effect.
PEG
44. • Artificial phospholipid vesicles
• Lipofection.
• Fraley in 1988
• DNA complex with liposomes
• Liposome +ve charage
• Membrane – membrane fusion
• Endocytosis
Liposome
46. • Advantages
–Simplicity
–Long term stability
–Low toxicity
–Protection of nucleic acid from degradation
• Limitations
–DNA passes from cytoplasm to the nucleus
unknown
Liposome
47. • Polycationic high molecular weight
• Endocytosis
• Along with DMSO and PMMA (Polymethyl-
methacrylate) - high
• Viral infection in cell lines.
• 80% transformed can express.
• Transfer efficiency increased by Glycerol.
DEAE Dextran
50. • Zimmermann et al., 1983
• Electrical shock (Charge)
• Suspending solution
• Electroinjection
• Types
–High voltage short duration
–Low voltage long duration
Electroporation
51. • Advantages
–Method is applicable to variety of cell lines
–Method quick
–Large number cells handle simultaneously
–Less cost
• Limitations
–Degradation of DNA
–Cell structure
–Clumping of cell
Electroporation
52.
53. • Mostly protoplast
• Mixed fusion
• Two types fusion (Cybrid, Hybrid)
• Along with PEG
• Sensitive to high PEG
Electrofusion
55. • Advantages
–Fusion very easy
–Fuse more cells
• Limitations
–Degradation of DNA
–Cell structure
–Clumping of cell
–Integrated DNA may be modified
Electrofusion