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    Question 5

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    Question 5

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    5 views2 pages

    Question 5

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    integral students tube

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    5.

    Explain the technique of rDNA to produce large amount of B cells


    from their source (1 page)
    The technique of recombinant DNA (rDNA) is widely used in biotechnology to produce large amounts of
    specific proteins, including B cells. The process involves combining DNA from different sources to create
    a novel DNA sequence that can be expressed in a host organism, such as bacteria or mammalian cells,
    leading to the production of the desired protein.

    To produce large amounts of B cells using rDNA technology, the following steps are typically followed:

     Identification of the target B cell protein: The first step is to identify the specific B cell protein of
    interest. This could be an antibody or any other protein produced by B cells. The DNA sequence
    encoding this protein needs to be known or isolated from the source organism.
     Isolation of the DNA sequence encoding the target protein: The DNA sequence encoding the
    target protein is isolated from the genome of the source organism. This can be done using
    techniques like polymerase chain reaction (PCR) or restriction enzymes. Once isolated, the DNA
    sequence is purified and ready for further manipulation.
     Cloning the DNA sequence into a suitable vector: A vector, typically a plasmid, is used to
    facilitate the insertion and expression of the target DNA sequence into a host organism. The
    isolated DNA sequence is ligated into the vector using DNA ligase, resulting in a recombinant
    DNA molecule.
     Transformation of the host organism: The recombinant DNA molecule is introduced into a host
    organism. Common host organisms used for rDNA technology include bacteria, yeast, and
    mammalian cells. The choice of host organism depends on factors like protein folding
    requirements, post-translational modifications, and protein functionality.
     Selection and amplification of transformed cells: In order to select and amplify cells that have
    taken up the recombinant DNA, a selectable marker is included in the vector. This marker allows
    for the identification and isolation of cells that have successfully incorporated the target DNA
    sequence. Commonly used markers include antibiotic resistance genes, fluorescent proteins, or
    enzymes for metabolic selection.
     Expression and purification of the target protein: The transformed host cells are grown in
    culture conditions that promote protein expression. The host cells then use their own machinery
    to transcribe the DNA into messenger RNA (mRNA) and translate it into the target protein.
    Various purification techniques, such as chromatography, are employed to separate and purify
    the protein of interest from other cellular components.
     Verification of protein functionality: Once the target protein has been purified, its functionality
    is verified using a range of assays, such as ELISA or Western blotting. These assays confirm that
    the recombinant B cells are producing the desired protein, which can then be used for various
    applications like research, diagnostics, or therapeutics.

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