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    Gene Cloning

    Uploaded by

    Indrakanth Krish
    Gene cloning allows for the synthesis of multiple copies of a particular DNA sequence. There are two main methods of gene cloning: in vivo and in vitro. The process involves amplifying a specific gene, cutting DNA at precise locations, joining DNA pieces, selecting replicating DNA, and transferring vectors into host cells. Plasmids are commonly used as cloning vectors since they can be modified to carry new genes and have origins of replication and antibiotic resistance markers. Applications of gene cloning include producing recombinant proteins like interferons, antibodies, and hormones for medical use as well as modifying crops for improved nutrition and insect resistance. However, there are also risks like low cloning success rates and potential for causing cancers.

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    Gene Cloning

    Uploaded by

    Indrakanth Krish
    1K views18 pages
    Gene cloning allows for the synthesis of multiple copies of a particular DNA sequence. There are two main methods of gene cloning: in vivo and in vitro. The process involves amplifying a specific gene, cutting DNA at precise locations, joining DNA pieces, selecting replicating DNA, and transferring vectors into host cells. Plasmids are commonly used as cloning vectors since they can be modified to carry new genes and have origins of replication and antibiotic resistance markers. Applications of gene cloning include producing recombinant proteins like interferons, antibodies, and hormones for medical use as well as modifying crops for improved nutrition and insect resistance. However, there are also risks like low cloning success rates and potential for causing cancers.

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    Gene cloning allows for the synthesis of multiple copies of a particular DNA sequence. There are two main methods of gene cloning: in vivo and in vitro. The process involves amplifying a specific gene, cutting DNA at precise locations, joining DNA pieces, selecting replicating DNA, and transferring vectors into host cells. Plasmids are commonly used as cloning vectors since they can be modified to carry new genes and have origins of replication and antibiotic resistance markers. Applications of gene cloning include producing recombinant proteins like interferons, antibodies, and hormones for medical use as well as modifying crops for improved nutrition and insect resistance. However, there are also risks like low cloning success rates and potential for causing cancers.

    Copyright:

    © All Rights Reserved
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    1K views18 pages

    Gene Cloning

    Uploaded by

    Indrakanth Krish
    Gene cloning allows for the synthesis of multiple copies of a particular DNA sequence. There are two main methods of gene cloning: in vivo and in vitro. The process involves amplifying a specific gene, cutting DNA at precise locations, joining DNA pieces, selecting replicating DNA, and transferring vectors into host cells. Plasmids are commonly used as cloning vectors since they can be modified to carry new genes and have origins of replication and antibiotic resistance markers. Applications of gene cloning include producing recombinant proteins like interferons, antibodies, and hormones for medical use as well as modifying crops for improved nutrition and insect resistance. However, there are also risks like low cloning success rates and potential for causing cancers.

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    Gene cloning

    By
    D.Dolabza
    V . s. lakshmi womens degree college
    What is gene cloning ?
    The process of synthesizing multiple copies of a particular DNA sequence
    using a bacteria cell or another organism as a host.
    GENE CLONING

    TWO METHODS
    1.) in vivo 2.) in vitro
    STEPS INVOLVED IN GENE CLONING
    1.) Amplification of specific gene

    2.) Cutting DNA at precise location

    3.) Join two pieces of DNA

    4.) Selection of small self replicating DNA

    5.) Method to move a vector into a host

    6.) Method to select host expressing recombinant DNA


    PLASMIDS – VEHICLES FOR CLONING

     Plasmids are naturally occurring Ampr

    extrachromosomal DNA molecules. Ori


    pBR322
     Plasmids are circular, double-stranded 4361bp

    DNA. Tetr

     Plasmids are the means by which


    antibiotic resistance is often transferred
    from one bacteria to another.
     Plasmids can be cleaved by restriction LacZ
    enzymes, leaving sticky or blunt ends. MCS

     Artificial plasmids can be constructed by pUC18


    Ori
    linking new DNA fragments to the sticky
    Ampr
    ends of plasmid.
    CLONING VECTORS Older cloning vector

     A cloning vector is a plasmid that can be Ampr

    modified to carry new genes. Ori


    pBR322
    4361bp
     Plasmids useful as cloning vectors must have:
    • An origin of replication. Tetr

    • A selectable marker (antibiotic resistance

    gene, such as amp1 and tetr).


    Newer cloning vector
    • Multiple cloning site (MCS) (site where

    insertion of foreign DNA will not disrupt LacZ


    MCS

    replication or inactivate essential


    pUC18
    markers). Ori

    • Easy to purify away from host DNA. Ampr


    Applications of gene cloning
    Medical utility :-
    Interferons Antibiotics

    Vitamins Harmones
    Interferons
     Natural proteins produced by the cells of the immune system
    of most vertebrates in response to challenges by foreign
    agents such as viruses, parasites and tumor cells.

     Interferons belong to the large class of glycoproteins


    known as cytokines.
    Interferons assist the immune response by :
    1. Inhibiting viral replication within host cells,

    2.Activating natural killer cells and macrophage

    3.Increasing antigen presentation to lymphocytes,

    4.Inducing the resistance of host cells to viral infectio


    Some recombinant DNA products being used in
    human therapy
    INSULIN for diabetics
    HUMAN GROWTH HARMONES (HGH)

    ERYTHROPOIETIN (EPO) for treating anemia


    Several types of INTERFERONS

    Many MONOCLONAL ANTIBODIES


    Agriculture utility :-

    Bacillus thuringiensis bacteria produce a protein toxin that kills insect larvae
    pests and is 80,000 times more toxic than the typical chemical insecticide.

    Transgenic tomato, corn, potato, and cotton plants have been made that produce a
    toxin from B. thuringiensis.
    Grains with improved nutritional characteristics:
    Genes from bacteria and daffodil plants are transferred to rice using the
    vector
    Agrobacterium tumefaciens

    Now a genetically modified strain of rice produces b-carotene, a


    molecule that to vitamin A in animals.
    is converted
    Attempts for impossible tasks

     Cleaning of :DDT

    Industrial wastes

    Reduce viscosity of crude oils

    Conversion of sunlight to electricity

    Degrade sulphur to coal

    Bacterial fermentation to produce methane and other energy forms

    Plastic degradation
    Biodegradable plastic-degrading enzyme from Pseudozyma antarcti

    Pseudozyma antarctica JCM 10317 exhibits a strong degradation activity for biodegradable
    plastics (BPs) such as agricultural mulch films composed of poly( butylene succinate) (PBS)
    and poly(butylene succinate-co-adipate) (PBSA).

    An enzyme named PaE was isolated and the gene encoding PaE was cloned from
    the strain by functional complementation in Saccharomyces cerevisiae.
    GLOWING CACTUS
    Risks of cloning

     >90% of cloning attempts fail to produce viable


    offsprings
    Cancers are caused

    Higher rates of infections


    Conclusion
    Thank you
    For
    Your Attention
    A
    PPT by
    Dolabza

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