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  • Gene Transfer

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    Ashima Goel
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    Gene transfer is the process of transferring a gene from one DNA molecule to another. There are several mechanisms for gene transfer including conjugation, bacterial transformation, retroviral transduction, and Agrobacterium-mediated transfer. Microinjection is a novel and efficient gene transfer technique that directly delivers DNA inside living cells using microscopic devices like glass micropipettes. The DNA is injected into the cell where it can penetrate the cell membrane and nuclear envelope to transfer the desirable gene. If the transfer is successful, the transformed cell can be cultured to develop into a transgenic plant. Biolistics, or particle bombardment, is another effective method for gene transfer where DNA is coated onto microprojectiles that are accelerated into plant tissues using a

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    Gene transfer is the process of transferring a gene from one DNA molecule to another. There are several mechanisms for gene transfer including conjugation, bacterial transformation, retroviral transduction, and Agrobacterium-mediated transfer. Microinjection is a novel and efficient gene transfer technique that directly delivers DNA inside living cells using microscopic devices like glass micropipettes. The DNA is injected into the cell where it can penetrate the cell membrane and nuclear envelope to transfer the desirable gene. If the transfer is successful, the transformed cell can be cultured to develop into a transgenic plant. Biolistics, or particle bombardment, is another effective method for gene transfer where DNA is coated onto microprojectiles that are accelerated into plant tissues using a

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    21 views12 pages

    Gene Transfer

    Uploaded by

    Ashima Goel
    Gene transfer is the process of transferring a gene from one DNA molecule to another. There are several mechanisms for gene transfer including conjugation, bacterial transformation, retroviral transduction, and Agrobacterium-mediated transfer. Microinjection is a novel and efficient gene transfer technique that directly delivers DNA inside living cells using microscopic devices like glass micropipettes. The DNA is injected into the cell where it can penetrate the cell membrane and nuclear envelope to transfer the desirable gene. If the transfer is successful, the transformed cell can be cultured to develop into a transgenic plant. Biolistics, or particle bombardment, is another effective method for gene transfer where DNA is coated onto microprojectiles that are accelerated into plant tissues using a

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    © All Rights Reserved
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    of 12

    •Gene transfer is to transfer a gene from one

    DNA molecule to another DNA molecule


    Mechanism -> DNA transferred from donor to
    recipient

    recombinant cell

    •Genetic transformation Transient


    Stable

    •Transferred gene – transgene


    •Organism that develops after successful gene
    transfer - transgenic
    1. Conjugation • PHYSICAL
    2. Bacterial 1. Microinjection
    transformation 2. Biolistics transformation
    3. Retroviral • Chemical
    transduction 1. Calcium Phosphate method
    4. Agrobacterium 2. Liposome mediated transfer
    mediated transfer
    • Electrical
    1. Electroporation
    • Novel & effecient technique
    • DNA delivered directly inside living cell
    (intact cell/ protoplast)
    • Uses microcapillaries and microscopic
    devices
    • 2 types of pipettes
    1. Glass micropipette to deliver DNA (0.5-10
    um)
    2. Holding pipette
    • Exogenous DNA taken in micropipette/
    microinjector -> easily penetrates cell
    membrane and nuclear envelope ->
    transfers the desirable DNA
    • Cell / protoplast -> firmly immobilized
    by holding pipette which exerts suction
    pressure
    • As the process of microinjection is
    complete, the transformed cell is
    cultured and grown to develop into a
    transgenic plant. In fact, transgenic
    tobacco and Brassica napus have been
    developed by this approach.

    Microinjection
    into the cell of a
    potato callus.
    • Thequantity of DNA delivery can be optimized
    •Even a very small cell can be targeted to deliver
    DNA
    •Frequency of stable integration of DNA is far
    better as compared to other methods

    •Costly instruments
    •Requires skilled persons for microinjection
    •Only one cell receives DNA per injection
    Most effective method for gene transfer, and creation of transgenic
    plants
    Predominently used for plant transformations
     plant tissues , organs , even pollen can be transformed very
    efficiently
    initially named as biolistics ( inventor Sanford (1988) )
    Biolistics = Biological + Ballistics
    Other names ->  particle gun, gene gun, bio blaster
    • 2 types of projectile systems – macro and
    microprojectile (DNA seq. of interest coated on gold
    or tungsten particles of 1-3um)
    Video on gene gun
    Direct delivery of DNA in the nucleus
    One shot can lead to multiple hits i.e. transfer of
    genes into many cells
    Speedy and accurate result
    Transformed plant organs display efficient
    regeneration
    BT maize is a product of biollistics
     Plastid transformation 

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