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    Cointegrated Vectors

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    devikamurugan124206
    Co-integrated vectors are constructed by recombining a bacterial plasmid containing exogenous DNA with the T-DNA region of an Agrobacterium Ti plasmid. This requires three vectors: 1) a disarmed Ti plasmid with the oncogenes replaced, 2) an intermediate vector containing the exogenous T-DNA, and 3) a helper vector providing transfer genes. The resulting co-integrated plasmid contains vir genes, T-DNA borders, exogenous DNA, and selectable markers but is less commonly used than binary vectors due to requiring long homologous regions and being less efficient than binary vectors.

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    Cointegrated Vectors

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    devikamurugan124206
    628 views2 pages
    Co-integrated vectors are constructed by recombining a bacterial plasmid containing exogenous DNA with the T-DNA region of an Agrobacterium Ti plasmid. This requires three vectors: 1) a disarmed Ti plasmid with the oncogenes replaced, 2) an intermediate vector containing the exogenous T-DNA, and 3) a helper vector providing transfer genes. The resulting co-integrated plasmid contains vir genes, T-DNA borders, exogenous DNA, and selectable markers but is less commonly used than binary vectors due to requiring long homologous regions and being less efficient than binary vectors.

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    Co-integrated vectors are constructed by recombining a bacterial plasmid containing exogenous DNA with the T-DNA region of an Agrobacterium Ti plasmid. This requires three vectors: 1) a disarmed Ti plasmid with the oncogenes replaced, 2) an intermediate vector containing the exogenous T-DNA, and 3) a helper vector providing transfer genes. The resulting co-integrated plasmid contains vir genes, T-DNA borders, exogenous DNA, and selectable markers but is less commonly used than binary vectors due to requiring long homologous regions and being less efficient than binary vectors.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    628 views2 pages

    Cointegrated Vectors

    Uploaded by

    devikamurugan124206
    Co-integrated vectors are constructed by recombining a bacterial plasmid containing exogenous DNA with the T-DNA region of an Agrobacterium Ti plasmid. This requires three vectors: 1) a disarmed Ti plasmid with the oncogenes replaced, 2) an intermediate vector containing the exogenous T-DNA, and 3) a helper vector providing transfer genes. The resulting co-integrated plasmid contains vir genes, T-DNA borders, exogenous DNA, and selectable markers but is less commonly used than binary vectors due to requiring long homologous regions and being less efficient than binary vectors.

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    Co-integrated Vectors

    Called co-integrated vectors or hybrid Ti plasmids, these vectors were among the first types of modified and engineered Ti plasmids devised for Agrobacterium -mediated transformation, but are not widely used today. These vectors are constructed by homologous recombination of a bacterial plasmid with the T-DNA region of an endogenous Ti plasmid in Agrobacterium. Integration of the two plasmids requires a region of homology present in both. Three vectors are necessary in this system:

    Disarmed Agrobacterium Ti plasmids In these Ti plasmids, the oncogenes located in the T-DNA region have been replaced by exogenous DNA. Examples of these vectors include: 1. SEV series: the right border of the T-DNA together with the phytohormone genes coding for cytokinin and auxin are removed and replaced by a bacterial kanamycin resistance gene while the left border and a small part of the left segment (TL) of the original T-DNA (referred to as Left Inside Homology (LIH)) are left intact. 2. pGV series: the phytohormone genes are excised and substituted by part of pBR322 vector sequence. The left and right border sequences as well as the nopaline synthase gene of the Ti plasmid are conserved.

    Intermediate vectors These are small pBR322-based plasmids (E. coli vectors) containing a TDNA region. They are used to overcome the problems derived from the large size of disarmed Ti plasmids and their lack of unique restriction sites. Intermediate vectors are replicated in E.coli and are transferred into Agrobacterium by conjugation. They cannot replicate in A.

    tumefaciens and therefore, carry DNA segments homologous to the


    disarmed T-DNA to permit recombination to form a co-integrated T-DNA structure.

    Helper vectors These are small plasmids maintained in E. coli that contain transfer (tra) and mobilization (mob) genes, which allow the transfer of the conjugation-deficient intermediate vectors into Agrobacterium.

    A resulting co-integrated plasmid assembled by in vitro manipulation normally contains: 1. the vir genes, 2. the left and right T-DNA borders, 3. an exogenous DNA sequence between the two T-DNA borders, and 4. plant and bacterial selectable markers. Some drawbacks Although co-integrated vectors have been designed to allow site-specific recombination based on the recombination system of the phage P1 (e.g., wP1loxP-Cre series), co-integrated vectors in general are less popular due to:

    long homologies required between the Ti plasmid and the E.

    coli plasmids making them difficult to engineer and use, and


    relatively inefficient gene transfer compared to the binary vectors.

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