Plant Transformation Methods
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“It first has to enter the plant cell by penetrating the plant cell wall and the plasma membrane and then must reach the nucleus and integrate into the resident chromosomes.”
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This document summarizes the process of plant genetic transformation using Agrobacterium tumefaciens. It describes how A. tumefaciens transfers T-DNA from its Ti plasmid into plant cells, integrating the T-DNA into the plant genome and expressing genes that cause crown gall disease. The document also outlines the key steps in the process, from gene transfer to the plant cell through regeneration of a transformed whole plant and methods to detect successful transformation events. Common genes inserted into transgenic crops are also listed, including genes for herbicide and insect resistance.
Binary Vector, By KK Sahu sir
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
Feature of ti plasmid
It is a part of Ti Plasmid which takes part intransfer and integration of T-DNA into plant chromosome.
The vir sequence consist of 8 operons which take part in different functions associated with virulence of Ti Plasmid. These are vir H, vir A, vir B, vir G, vir C, vir D, vir E, & vir F. ( vir H & vir F present occasionally).
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This document discusses genetic engineering techniques using Agrobacterium tumefaciens and its tumor-inducing (Ti) plasmid. The Ti plasmid contains a segment of transferred DNA (T-DNA) that is integrated into the host plant genome and causes tumor formation. Genes of interest can be inserted into the T-DNA, which is then transferred to the plant cell. However, wild-type Ti plasmids also contain oncogenes, so alternative vectors like plant viruses have been developed that can introduce foreign DNA into plant cells without causing tumors. Cauliflower mosaic virus is one such virus vector that has been used, as its small, 8kb double-stranded DNA genome can be easily manipulated.
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Agrobacterium mediated gene transfer
This document summarizes the process of plant genetic transformation using Agrobacterium tumefaciens. It describes how A. tumefaciens transfers T-DNA from its Ti plasmid into plant cells, integrating the T-DNA into the plant genome and expressing genes that cause crown gall disease. The document also outlines the key steps in the process, from gene transfer to the plant cell through regeneration of a transformed whole plant and methods to detect successful transformation events. Common genes inserted into transgenic crops are also listed, including genes for herbicide and insect resistance.
Binary Vector, By KK Sahu sir
Introduction
Components of binary vector
Development of binary vector system
Properties of binary vector
Types of binary vector
Plant transformation using binary vector
Advantage of using binary vector
Conclusion
References
Feature of ti plasmid
It is a part of Ti Plasmid which takes part intransfer and integration of T-DNA into plant chromosome.
The vir sequence consist of 8 operons which take part in different functions associated with virulence of Ti Plasmid. These are vir H, vir A, vir B, vir G, vir C, vir D, vir E, & vir F. ( vir H & vir F present occasionally).
Ti plasmid and ca mv
This document discusses genetic engineering techniques using Agrobacterium tumefaciens and its tumor-inducing (Ti) plasmid. The Ti plasmid contains a segment of transferred DNA (T-DNA) that is integrated into the host plant genome and causes tumor formation. Genes of interest can be inserted into the T-DNA, which is then transferred to the plant cell. However, wild-type Ti plasmids also contain oncogenes, so alternative vectors like plant viruses have been developed that can introduce foreign DNA into plant cells without causing tumors. Cauliflower mosaic virus is one such virus vector that has been used, as its small, 8kb double-stranded DNA genome can be easily manipulated.
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description of Ti plasmid derivatives as vector and explanation of T DNA and cointegrate vectors and binary vectors and its uses and advantages and disadvantages
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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.
Plant transformation vectors and their types
This document summarizes a presentation on plant transformation vectors and their types. It discusses various types of vectors used for plant transformation including plasmids, viruses, bacteriophages, and cosmids. Plasmids are the most commonly used vector for plant transformation. Agrobacterium-mediated transformation using tumor-inducing (Ti) plasmids is an effective method for genetically transforming plants. Viral vectors like cauliflower mosaic virus (CaMV) are also used but have certain limitations.
Cauliflower mosaic virus ppt
INTRODUCTION:
The first plant virus shown to have a DNA genome and the first shown to replicate by reverse transcription.
Worldwide but only causes significantly losses locally.
It is transmitted by aphids .
Type member of the Caulimovirus genus, contains 11 species and 6 possible members.
significantly impact on plant virology and plant molecular biology.
The virus is an important source of gene regulatory elements, used exclusively in the genetic manipulation of plants.
STRUCTURE:Icosachedral with a diameter of 52Â nm built from 420 capsid protein subunits.
It contains a circular double-stranded DNA molecule of about 8.0 kB .
Dna is interrupted by sitespecific discontinuties resulting from its replication by reverse transcription.
After entering the host, the single stranded nicks in the viral DNA are repaired, forming a supercoiled molecule that binds to histones.
DNA is transcriped into a full length .
Replication
Risk Factors:The Cauliflower mosaic virus promoter (CaMV 35S) is used in most transgenic crops to activate foreign genes which have been artificially inserted into the host plant. It is inserted into transgenic plants in a form which is different from that found when it is present in its natural Brassica plant hosts. This enables it to operate in a wide range of host-organism environments which would otherwise not be possible.
vector and its types
Vector and its types.
@ Zaheer
International Islamic University Islamabad.
Mail id. muhammad.bsbt642@iiu.edu.pk
cloning and expression vector in plants
This document discusses cloning and expression vectors used in plants. It describes cloning vectors as small DNA molecules that can stably maintain foreign DNA for cloning purposes. Expression vectors are designed for protein expression and must contain strong promoters and terminators. Common vectors used in plants include plasmids, viruses, bacteriophages, and cosmids. Agrobacterium-mediated transformation is described as a method for introducing DNA into plants using Ti or Ri plasmids. Viral vectors like CaMV can also be used to directly or indirectly insert genes into plants.
DNA containing plant viruses - Cauliflower mosaic virus & Geminivirus
This document provides a summary of a seminar on DNA containing plant viruses. It discusses the Cauliflower Mosaic Virus (CaMV) and Gemini virus. CaMV is a double-stranded DNA virus that infects members of the Brassicaceae family and causes mosaic symptoms. It replicates via reverse transcription. Gemini virus is a single-stranded DNA virus that infects many economically important crops and causes stunting and yellowing. It replicates via a rolling circle mechanism and is transmitted by insects like leafhoppers and whiteflies. Prevention of both viruses focuses on controlling their insect vectors and disinfecting equipment to avoid mechanical transmission.
agrobacterim vector
The document summarizes how Agrobacterium tumefaciens is used for plant genetic engineering. It describes the key components of the Ti plasmid, including the T-DNA, vir genes, and opine catabolism genes. It explains that the Ti plasmid is modified by deleting tumor-causing genes and inserting a gene of interest, along with selectable markers. The modified plasmid is transferred to A. tumefaciens via triparental mating and then used to transform plant cells. Transformed cells are selected using antibiotics and regenerated into transgenic plants.
Vectors Used for Gene Cloning in Plants
Cloning vectors are small DNA molecules that are used to artificially carry foreign genetic material into host cells. They contain features like an origin of replication, antibiotic resistance genes, and restriction enzyme sites. The document discusses different types of cloning vectors used for plant gene cloning, including plasmids, Ti/Ri plasmids from Agrobacterium, and plant viruses. Agrobacterium-mediated transformation uses disarmed Ti plasmids from Agrobacterium tumefaciens and involves co-cultivation of plant explants with Agrobacterium, selection of transformed cells, and regeneration of whole plants. Binary vector systems are now commonly used, involving transfer of a binary plasmid without integration into the Ti plasmid
Ti plasmid as a vector,
Introduction
Ti plasmid
Agrobacterium tumefaciens
Ti plasmid structure
Overview of infection process
Ti plasmid derived vector systems
Cointegrate vectors
Binary vectors
Agrobacterium mediated transformation of explants
Conclusions
References
Cauliflower mosaic virus
CaMV Genome organization & their replication, Cauliflower Mosaic Virus belong to Group VII (ds-DNA-RT), Open circular double stranded DNA of 80kb and CaMV replicates by reverse transcription
Agrobacterium mediated gene transfer
The life cycle of Agrobacterium species in a plant and its infection process. This natural mechanism of gene transfer can be manipulated and put for use by geneticists to transfer foreign genes of interest.
Agrobacterium mediated gene transfer in plants.
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.
Microinjection ssb
Microinjection is a technique for introducing DNA into plant cells by directly injecting it through the cell membrane using a microscopic needle. The document describes microinjection and its application in crop improvement. It details the microinjection process, including preparing the needle and holding the cell, precisely injecting the DNA, and recovering transgenic plants. Advantages are precise delivery and ability to introduce various DNA structures. Disadvantages include being costly, limited to protoplasts, and slow. As a case study, cotton plants were transformed via microinjecting a modified cry1Ia gene into flower ovaries, resulting in a transgenic cotton line with the full cry1Ia gene integrated into its genome.
Cauliflower mosaic virus
Cauliflower Mosaic Virus is a pararetrovirus that infects plants in the brassicaceae family like cauliflower. It has an icosahedral capsid containing a circular double stranded DNA genome around 80kb in size. The virus replicates through reverse transcription, with its DNA entering the nucleus and being transcribed by the host polymerase. The virus has several open reading frames that encode for structural, movement and other proteins. While it can be used as a vector to insert foreign genes into plants, its capacity is limited to a few hundred nucleotides before the foreign DNA is expelled.
Applications of Genetic Engineering
This presentation gives an brief idea about the applications of genetic engineering which is of at most importance to humans. Provided along with this slide is an example which makes it easier to understand the concept.
Sv40 virus
Simian virus 40 (SV40) is a polyomavirus that was first isolated from monkey kidney cell cultures used to produce the poliovirus vaccine. It has a small circular DNA genome wrapped in histone proteins. The genome contains early and late protein coding genes as well as regulatory sequences. The early proteins are non-structural while the late proteins form the viral capsid. The SV40 genome can be modified to replace the early protein coding genes with a gene of interest for gene therapy purposes. A recombinant SV40 viral vector offers advantages such as the ability to transduce both dividing and resting cells with high efficiency and provide long-lasting transgene expression.
Intracellular highways in the plants: the role of the cytoskeleton in camv i...
The document discusses research on the intracellular movement of Cauliflower Mosaic Virus (CaMV) particles. It finds that the CaMV P6 protein forms motile inclusion bodies that traffic along the plant cell's actin microfilaments and stabilize microtubules. The P6 inclusion bodies are hypothesized to function as "virion factories" where CaMV particles assemble before being transported to plasmodesmata for movement between cells. A yeast two-hybrid screen identified the host protein CHUP1, which mediates chloroplast movement along microfilaments, as interacting with P6. Silencing CHUP1 reduced the rate of CaMV lesion development, supporting its role in P6 inclusion body movement.
MICROINJECTION GENE TRANSFER METHOD
Microinjection is a gene transfer technique where DNA is directly injected into cells using a fine glass micropipette. It is highly efficient at the individual cell level and was originally used for transfecting hard-to-transfect cells. The procedure involves holding a cell using one pipette while another pipette is used to inject DNA into the cell's cytoplasm or nucleus. It allows for stable transfection efficiencies of around 20% and is used to generate transgenic animals by injecting DNA into oocytes, eggs or embryos. However, it is time-consuming and can only be done for a small number of cells.
Viral vectors in virology
definition,converting a virus into a vector,main types og viral vectors,principle of retroviral gene transfer,
L7. artificial chromosomes, plant viral vectors and animal viral vectors
This document discusses various cloning vectors that have been developed over time for use in genetic engineering, including plasmids, bacteriophages, cosmids, yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), P1 artificial chromosomes (PACs), and human artificial chromosomes (HACs). YACs were developed in 1987 but have issues with chimerism and instability. BACs were developed in 1992 and overcome many of the limitations of YACs. PACs were developed in 1994 and combine features of P1 vectors and BACs. HACs were first constructed in 1997 and provide a system for gene expression studies in human cells.
Ti Plasmid
Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into the genome of plant cells. Scientists have harnessed this natural ability of A. tumefaciens by modifying its Ti plasmid to replace the tumor-causing genes with a gene of interest. This disarmed Ti plasmid, along with helper plasmids containing virulence genes, allows for the stable integration and expression of the new gene in plant cells. Common transformation methods using A. tumefaciens involve culturing plant explants like leaf disks with Agrobacteria containing the modified Ti plasmid, then regenerating transgenic plants from the transformed cells
Agrobacterium mediated gene transfer
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.
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Ti plasmid
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.
Plant transformation vectors and their types
This document summarizes a presentation on plant transformation vectors and their types. It discusses various types of vectors used for plant transformation including plasmids, viruses, bacteriophages, and cosmids. Plasmids are the most commonly used vector for plant transformation. Agrobacterium-mediated transformation using tumor-inducing (Ti) plasmids is an effective method for genetically transforming plants. Viral vectors like cauliflower mosaic virus (CaMV) are also used but have certain limitations.
Cauliflower mosaic virus ppt
INTRODUCTION:
The first plant virus shown to have a DNA genome and the first shown to replicate by reverse transcription.
Worldwide but only causes significantly losses locally.
It is transmitted by aphids .
Type member of the Caulimovirus genus, contains 11 species and 6 possible members.
significantly impact on plant virology and plant molecular biology.
The virus is an important source of gene regulatory elements, used exclusively in the genetic manipulation of plants.
STRUCTURE:Icosachedral with a diameter of 52Â nm built from 420 capsid protein subunits.
It contains a circular double-stranded DNA molecule of about 8.0 kB .
Dna is interrupted by sitespecific discontinuties resulting from its replication by reverse transcription.
After entering the host, the single stranded nicks in the viral DNA are repaired, forming a supercoiled molecule that binds to histones.
DNA is transcriped into a full length .
Replication
Risk Factors:The Cauliflower mosaic virus promoter (CaMV 35S) is used in most transgenic crops to activate foreign genes which have been artificially inserted into the host plant. It is inserted into transgenic plants in a form which is different from that found when it is present in its natural Brassica plant hosts. This enables it to operate in a wide range of host-organism environments which would otherwise not be possible.
vector and its types
Vector and its types.
@ Zaheer
International Islamic University Islamabad.
Mail id. muhammad.bsbt642@iiu.edu.pk
cloning and expression vector in plants
This document discusses cloning and expression vectors used in plants. It describes cloning vectors as small DNA molecules that can stably maintain foreign DNA for cloning purposes. Expression vectors are designed for protein expression and must contain strong promoters and terminators. Common vectors used in plants include plasmids, viruses, bacteriophages, and cosmids. Agrobacterium-mediated transformation is described as a method for introducing DNA into plants using Ti or Ri plasmids. Viral vectors like CaMV can also be used to directly or indirectly insert genes into plants.
DNA containing plant viruses - Cauliflower mosaic virus & Geminivirus
This document provides a summary of a seminar on DNA containing plant viruses. It discusses the Cauliflower Mosaic Virus (CaMV) and Gemini virus. CaMV is a double-stranded DNA virus that infects members of the Brassicaceae family and causes mosaic symptoms. It replicates via reverse transcription. Gemini virus is a single-stranded DNA virus that infects many economically important crops and causes stunting and yellowing. It replicates via a rolling circle mechanism and is transmitted by insects like leafhoppers and whiteflies. Prevention of both viruses focuses on controlling their insect vectors and disinfecting equipment to avoid mechanical transmission.
agrobacterim vector
The document summarizes how Agrobacterium tumefaciens is used for plant genetic engineering. It describes the key components of the Ti plasmid, including the T-DNA, vir genes, and opine catabolism genes. It explains that the Ti plasmid is modified by deleting tumor-causing genes and inserting a gene of interest, along with selectable markers. The modified plasmid is transferred to A. tumefaciens via triparental mating and then used to transform plant cells. Transformed cells are selected using antibiotics and regenerated into transgenic plants.
Vectors Used for Gene Cloning in Plants
Cloning vectors are small DNA molecules that are used to artificially carry foreign genetic material into host cells. They contain features like an origin of replication, antibiotic resistance genes, and restriction enzyme sites. The document discusses different types of cloning vectors used for plant gene cloning, including plasmids, Ti/Ri plasmids from Agrobacterium, and plant viruses. Agrobacterium-mediated transformation uses disarmed Ti plasmids from Agrobacterium tumefaciens and involves co-cultivation of plant explants with Agrobacterium, selection of transformed cells, and regeneration of whole plants. Binary vector systems are now commonly used, involving transfer of a binary plasmid without integration into the Ti plasmid
Ti plasmid as a vector,
Introduction
Ti plasmid
Agrobacterium tumefaciens
Ti plasmid structure
Overview of infection process
Ti plasmid derived vector systems
Cointegrate vectors
Binary vectors
Agrobacterium mediated transformation of explants
Conclusions
References
Cauliflower mosaic virus
CaMV Genome organization & their replication, Cauliflower Mosaic Virus belong to Group VII (ds-DNA-RT), Open circular double stranded DNA of 80kb and CaMV replicates by reverse transcription
Agrobacterium mediated gene transfer
The life cycle of Agrobacterium species in a plant and its infection process. This natural mechanism of gene transfer can be manipulated and put for use by geneticists to transfer foreign genes of interest.
Agrobacterium mediated gene transfer in plants.
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.
Microinjection ssb
Microinjection is a technique for introducing DNA into plant cells by directly injecting it through the cell membrane using a microscopic needle. The document describes microinjection and its application in crop improvement. It details the microinjection process, including preparing the needle and holding the cell, precisely injecting the DNA, and recovering transgenic plants. Advantages are precise delivery and ability to introduce various DNA structures. Disadvantages include being costly, limited to protoplasts, and slow. As a case study, cotton plants were transformed via microinjecting a modified cry1Ia gene into flower ovaries, resulting in a transgenic cotton line with the full cry1Ia gene integrated into its genome.
Cauliflower mosaic virus
Cauliflower Mosaic Virus is a pararetrovirus that infects plants in the brassicaceae family like cauliflower. It has an icosahedral capsid containing a circular double stranded DNA genome around 80kb in size. The virus replicates through reverse transcription, with its DNA entering the nucleus and being transcribed by the host polymerase. The virus has several open reading frames that encode for structural, movement and other proteins. While it can be used as a vector to insert foreign genes into plants, its capacity is limited to a few hundred nucleotides before the foreign DNA is expelled.
Applications of Genetic Engineering
This presentation gives an brief idea about the applications of genetic engineering which is of at most importance to humans. Provided along with this slide is an example which makes it easier to understand the concept.
Sv40 virus
Simian virus 40 (SV40) is a polyomavirus that was first isolated from monkey kidney cell cultures used to produce the poliovirus vaccine. It has a small circular DNA genome wrapped in histone proteins. The genome contains early and late protein coding genes as well as regulatory sequences. The early proteins are non-structural while the late proteins form the viral capsid. The SV40 genome can be modified to replace the early protein coding genes with a gene of interest for gene therapy purposes. A recombinant SV40 viral vector offers advantages such as the ability to transduce both dividing and resting cells with high efficiency and provide long-lasting transgene expression.
Intracellular highways in the plants: the role of the cytoskeleton in camv i...
The document discusses research on the intracellular movement of Cauliflower Mosaic Virus (CaMV) particles. It finds that the CaMV P6 protein forms motile inclusion bodies that traffic along the plant cell's actin microfilaments and stabilize microtubules. The P6 inclusion bodies are hypothesized to function as "virion factories" where CaMV particles assemble before being transported to plasmodesmata for movement between cells. A yeast two-hybrid screen identified the host protein CHUP1, which mediates chloroplast movement along microfilaments, as interacting with P6. Silencing CHUP1 reduced the rate of CaMV lesion development, supporting its role in P6 inclusion body movement.
MICROINJECTION GENE TRANSFER METHOD
Microinjection is a gene transfer technique where DNA is directly injected into cells using a fine glass micropipette. It is highly efficient at the individual cell level and was originally used for transfecting hard-to-transfect cells. The procedure involves holding a cell using one pipette while another pipette is used to inject DNA into the cell's cytoplasm or nucleus. It allows for stable transfection efficiencies of around 20% and is used to generate transgenic animals by injecting DNA into oocytes, eggs or embryos. However, it is time-consuming and can only be done for a small number of cells.
Viral vectors in virology
definition,converting a virus into a vector,main types og viral vectors,principle of retroviral gene transfer,
L7. artificial chromosomes, plant viral vectors and animal viral vectors
This document discusses various cloning vectors that have been developed over time for use in genetic engineering, including plasmids, bacteriophages, cosmids, yeast artificial chromosomes (YACs), bacterial artificial chromosomes (BACs), P1 artificial chromosomes (PACs), and human artificial chromosomes (HACs). YACs were developed in 1987 but have issues with chimerism and instability. BACs were developed in 1992 and overcome many of the limitations of YACs. PACs were developed in 1994 and combine features of P1 vectors and BACs. HACs were first constructed in 1997 and provide a system for gene expression studies in human cells.
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Similar to Plant Transformation Methods
Ti Plasmid
Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments called T-DNA from its tumor-inducing (Ti) plasmid into the genome of plant cells. Scientists have harnessed this natural ability of A. tumefaciens by modifying its Ti plasmid to replace the tumor-causing genes with a gene of interest. This disarmed Ti plasmid, along with helper plasmids containing virulence genes, allows for the stable integration and expression of the new gene in plant cells. Common transformation methods using A. tumefaciens involve culturing plant explants like leaf disks with Agrobacteria containing the modified Ti plasmid, then regenerating transgenic plants from the transformed cells
Agrobacterium mediated gene transfer
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.
Plant Genetic Engineering & Agrobacterium mediated gene transfer.pptx
Agrobacterium mediated gene transfer, Ti-plasmid, cloning vectors based on Ti-plasmid, advantages disadvantages regarding cloning vectors based on Ti-plasmid are major areas covered in this Presentation.
This bacterium has a large plasmid that induces tumor, and for this reason, it was named tumor-inducing (Ti) plasmid.
This is process of altering the genetic makeup of an organism using Recombinant DNA Technology.
Plant transformation gene transfer methods in plants
The ultimate objective of modern plant breeding is to improve a top variety in one single additional character in a predictable and precise manner without disturbing the rest of the genome. Today this is being realised through examples of successful transfer of specific traits into higher plants by gene transfer.
Techniques that open up to the plant breeder the possibility of transferring in a planned manner characters from one organism to another have been developed in microbial genetics. It should be stressed right at the outset that the expression “gene” has different meanings in agriculture and in molecular biology.
Gene Transfer Methods:
The gene transfer techniques in plant genetic transformation are broadly grouped into two categories:
I. Vector-mediated gene transfer
II. Direct or vector less DNA transfer
Agrobacterium tumefaciens
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.
Biotechnology_transgenic plants _II.pptx
Genetically modified plants can be created through Agrobacterium-mediated gene transfer. Agrobacterium contains a Ti plasmid that can transfer DNA fragments into plant cells. Researchers have developed artificial Ti plasmids and binary vector systems to insert desirable genes. The binary system uses a disarmed helper Ti plasmid containing vir genes and a separate vector with the gene of interest. The cointegrate system allows the vector to recombine and integrate into the Ti plasmid. These systems allow Agrobacterium to transfer genes into plant genomes without causing tumors. Other gene transfer methods include particle bombardment and protoplast transformation. Molecular tests can identify transgenes in GM crops.
Agrobacterium mediated gene transfer in plants
1) Agrobacterium tumefaciens is a soil bacterium that causes crown gall disease in dicot plants by transferring oncogenic DNA (T-DNA) from its Ti plasmid into the plant genome.
2) The T-DNA contains genes that produce hormones causing uncontrolled cell division and the formation of tumors.
3) By removing the oncogenic genes and inserting foreign DNA between the border repeats, the Ti plasmid can be used as a vector to stably introduce foreign genes into plant genomes through the natural plant-Agrobacterium interaction and T-DNA transfer process.
Agrobacterium tumefacience
Agrobacterium tumefaciens is a soil bacterium that naturally transfers DNA to plant cells and causes crown gall disease. It contains a tumor-inducing plasmid (Ti plasmid) that can be engineered to transfer foreign genes of interest into plant genomes. This process, called Agrobacterium-mediated transformation, is commonly used in genetic engineering due to its high efficiency. The Ti plasmid contains T-DNA that is transferred to plant cells, along with virulence genes that facilitate transfer without being transferred themselves. Transformed plants are regenerated from cultured plant cells or tissues containing the novel genes. Agrobacterium-mediated transformation is useful for genetic engineering but has limitations such as a narrow host range and time-consuming
Gene transfer in plants
plant biotechnology, biotechnological tools, indirect gene transfer, vector, agrobacterium tumifaciens mediated gene transfer , Ti plasmid, T- DNA , grown gall disease
Agrobact transfer.pptx
Agrobacteria are soil bacteria that can transfer genes to plant genomes. They contain Ti plasmids which transfer a segment of DNA (T-DNA) into the host plant genome. This causes tumors and induces the plant to produce unique amino acids called opines that the bacteria can use as nutrients. The Ti plasmid and bacterial chromosome work together with virulence genes on the plasmid producing proteins needed for T-DNA transfer. The T-DNA is processed and transferred via a T-strand into the plant cell nucleus where it integrates randomly. This process allows genetic engineering of plants through Agrobacterium.
Ti plasmid ss
1. Agrobacterium tumefaciens is a soil bacteria that causes crown gall disease in plants by transferring a segment of DNA (T-DNA) from its tumor-inducing plasmid (Ti plasmid) into the plant genome.
2. The T-DNA contains oncogenes that cause tumor formation as well as genes for producing nutrients called opines that the bacteria can use.
3. Ti plasmids are modified to create disarmed vectors for plant transformation by deleting the oncogenes from the T-DNA. Binary vectors further separate the T-DNA and virulence genes into two plasmids.
Agrobacterium and other methods of plant transformation including gene gun, i...
The process of transfer, integration and expression of transgene in the host cells is known as genetic transformation. A foreign gene (transgene) encoding the trait must be incorporated into plant cells, along with a "cassette" of extra genetic material to add a desirable trait to a crop. The cassette includes a sequence of DNA called a "promoter", which determines where and when the foreign gene is expressed in the host, and a "marker gene" which allows breeders to determine by screening or selection which plants contain the inserted gene. For example, marker genes may make plants resistant to antibiotics not used routinely (e.g., agrimycin, kanamycin) or tolerant of some herbicides.
Biotechnology_transgenic plants.pptx
Genetically modified crops can be created through transgenic approaches using Agrobacterium tumefaciens. A. tumefaciens contains a tumor-inducing plasmid that allows it to transfer DNA fragments into plant cells. Researchers have developed methods to modify this plasmid to transfer desirable genes instead of tumor-causing ones. The modified plasmid is introduced into A. tumefaciens along with the gene of interest, and the bacteria is used to infect plant cells and transfer the new gene. This allows crops to be engineered for traits like pest resistance, herbicide tolerance, or improved nutrition more efficiently than traditional breeding methods.
Biological method of transformation
Agrobacterium and plant viruses can be used as biological vectors for plant transformation. Agrobacterium mediates transformation via its tumor-inducing plasmid, using virulence genes to transfer T-DNA containing the gene of interest into the plant genome. Plant viruses can also act as gene vectors by engineering viral genomes to contain and deliver foreign genes. Retroviruses have been developed as viral vectors for animal gene transfer due to their ability to stably integrate into the host chromosome.
Agrobacterium mediated transformation, its mode of action and applications in...
Agrobacterium mediated transformation, its mode of action and applications in...Dr. Shobha D. Surbhaiyya
here described in detail gene transformation technique in plant by natural method for improvement of crop Plant transformation methods
the presentation is provided with information about how the foreign genes are transferred to plant genome.
Agrobacterium mediated transformation
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.
Plant expression vectors
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.
Agrobacterium CM
1) Agrobacterium tumefaciens is a soil bacterium that can transfer DNA fragments (T-DNA) from its tumor-inducing plasmid into plant cells.
2) The T-DNA is flanked by left and right border sequences and encodes genes that cause plant cells to form tumors and produce nutrients for the bacteria.
3) Upon detection of wounded plant cells, genes on the bacterial plasmid and chromosome mediate T-DNA processing and transfer into the plant cell nucleus where it integrates randomly.
Agrobacterium - mediated Gene Transfer old.pptx
This document summarizes Agrobacterium-mediated plant genetic transformation. It describes how Agrobacterium transfers DNA (T-DNA) from its tumor-inducing (Ti) plasmid into the plant genome, causing crown gall disease. The T-DNA contains genes that cause plants to produce unique amino acids called opines, which the bacteria can use for food. Early transformation systems involved directly modifying the Ti plasmid, but this was limited. The binary vector system separates the T-DNA from virulence genes, allowing for easier genetic engineering. The co-integration strategy first clones the gene of interest into an E. coli vector before recombining into a Ti plasmid, but requires long homologous regions between plasmids.
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Plant Transformation Methods
- 2. Plant TransformationMethods “It first has to enter the plant cell by penetrating the plant cell wall and the plasma membrane and then must reach the nucleus and integrate into the resident chromosomes.” Agrobacterium tumefaciens. Particle bombardment.
- 3. The AgrobacteriumSystem A member of the eubacterial family Rhizobiaceae. The organism responsible for the elicitation of crown gall tumors in plants. formation of these Tumors occurs as a result of bacterial infection, usually at wound sites, on many dicotyledonous and some monocotyledonous plants,
- 4. The AgrobacteriumSystem Armin Braun demonstrated that tumor cells are transformed and that the uncontrolled proliferation of the tumor cells was not dependent on the continuous presence of Agrobacterium, implying the existence of a transformation-inducing principle.
- 5. The AgrobacteriumSystem Ivo Zaenen. Jeff Schell. Marc Van Montagu. virulent strains. X avirulent strains.
- 6. The AgrobacteriumSystem Eugene Nester. Milton Gordon. Mary-Dell Chilton.
- 8. The AgrobacteriumSystem heterologous genes with the appropriate regulatory regions into the T-DNA region and showed that foreign genes became integrated and functionally expressed in plant cells.
- 10. The AgrobacteriumSystem (a) In a significant percentage of the transformation events, a single copy of the T-DNA is integrated into the chromosomes of the transformed cell. (b) Numerous vector systems are now available containing the T- DNA borders and various reporter and selectable marker genes, allowing researchers to choose the most appropriate combination to insert heterologous genes. (c) It is possible to transfer large fragments of DNA, including bacterial artificial chromosomes. (d) Transformation in planta, without the necessity of tissue culture, is possible in some species such as Arabidopsis thaliana and Medicago trunculata
- 11. Transformationprocess (a) T-DNA border sequences, which demarcate the DNA segment (T-DNA) to be transferred into the plant genome. (b) vir genes (virulence genes), which are required for transferring the T-DNA region to the plant but are not themselves transferred. (c) modified T-DNA region where the genes that cause crown gall formation are removed and replaced with the genes of interest.
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