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    Recombinant DNA Technology

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    Amjad
    Recombinant DNA technology involves chemically joining together DNA molecules from different sources. The goals are to isolate, characterize, and alter genes, and return them to living cells to understand hereditary diseases, improve the human genome, and more. The procedure involves isolating DNA, cutting it with restriction enzymes, and joining the DNA with DNA ligase before amplifying the recombinant DNA in bacterial host cells. Various enzymes and vectors are used to replicate and study the recombinant DNA for applications in agriculture, medicine, pharmacology and more.

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    Recombinant DNA Technology

    Uploaded by

    Amjad
    152 views5 pages
    Recombinant DNA technology involves chemically joining together DNA molecules from different sources. The goals are to isolate, characterize, and alter genes, and return them to living cells to understand hereditary diseases, improve the human genome, and more. The procedure involves isolating DNA, cutting it with restriction enzymes, and joining the DNA with DNA ligase before amplifying the recombinant DNA in bacterial host cells. Various enzymes and vectors are used to replicate and study the recombinant DNA for applications in agriculture, medicine, pharmacology and more.

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    Recombinant DNA technology involves chemically joining together DNA molecules from different sources. The goals are to isolate, characterize, and alter genes, and return them to living cells to understand hereditary diseases, improve the human genome, and more. The procedure involves isolating DNA, cutting it with restriction enzymes, and joining the DNA with DNA ligase before amplifying the recombinant DNA in bacterial host cells. Various enzymes and vectors are used to replicate and study the recombinant DNA for applications in agriculture, medicine, pharmacology and more.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    152 views5 pages

    Recombinant DNA Technology

    Uploaded by

    Amjad
    Recombinant DNA technology involves chemically joining together DNA molecules from different sources. The goals are to isolate, characterize, and alter genes, and return them to living cells to understand hereditary diseases, improve the human genome, and more. The procedure involves isolating DNA, cutting it with restriction enzymes, and joining the DNA with DNA ligase before amplifying the recombinant DNA in bacterial host cells. Various enzymes and vectors are used to replicate and study the recombinant DNA for applications in agriculture, medicine, pharmacology and more.

    Copyright:

    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    of 5

    Recombinant DNA technology

    DNA molecules that are extracted from different sources and chemically joined together is called
    recombinant DNA and this technology is called recombinant DNA technology.

    Goals of recombinant DNA technology


    • To isolate and characterize a gene
    • To make desired alterations in one or more isolated genes
    • To return altered genes to living cells
    • Artificially synthesize new gene
    • Alternating the genome of an organism
    • Understanding the hereditary diseases and their cure
    • Improving human genome

    Procedure of making rDNA


    1. Isolating of DNA
    2. Cutting of DNA
    3. Joining of DNA
    4. Amplifying of DNA
    1. Isolating of DNA

    2. Cutting of DNA
    • DNA can be cut into large fragments by mechanical
    shearing.
    • Restriction enzymes are the scissors of molecular
    genetics.
    Restriction enzymes
    A special class of sequence-specific enzyme
    • Found in bacteria
    • Site-specific-cleave DNA molecules only at
    specific nucleotide sequence
    • REases recognize DNA base sequence that
    are palindrome
    • REase make staggered cuts with
    complementary base sequences for easy
    circulization
    3. Joing of DNA by DNA Ligase
    4. Amplifying the recombinant DNA
    • Transforming the recombinant DNA into a bacterial host strain.
    • The cells are treated with CaCl2
    • DNA is added
    • Cells are heat shocked at 42 C
    • DNA goes into cell by a somewhat unknown mechanism.
    • Once in a cell, the recombinant DNA will be replicated.
    • When the cell divides, the replicated recombinant molecules go to both
    daughter cells which themselves will divide later. Thus, the DNA is
    amplified

    Enzymes used in recombinant DNA technology


    DNA ligase: Bind to DNA molecules
    Type II restriction endonuclease: Cleaves DNA at specific sites
    Reverse transcriptase: Make a DNA copy of RNA molecule
    DNA polymerase I: Fill single stranded gapes of DNA duplex
    Polynycleotide Kinase: Adds a phosephate to the 5'-OH end of a polynucleotide
    Terminal transferase: Adds homopolymer tails to the 3'-OH ends
    Exonuclease III: Removes nucleotide residues from the 3' ends
    Bacteriophage {lamda} exonuclease: removes nucleotides from the 5' ends
    Alkaline phosphatase: Removes terminal phosphates

    Vectors used in rDNA technology


    • A vector is an area of DNA that can join another DNA
    part without losing the limit for self-replication
    • Should be capable of replicating in host cell
    • Should have convenient RE sites for inserting DNA of
    interest
    • Should have a selectable marker to indicate which
    host cells received recombinant DNA molecule
    • Should be small and easy to isolate

    Applications of rDNA technology


    • Agriculture: growing crops of your choice (GM food),
    pesticide resistant crops, fruits with attractive colors, all
    being grown in artificial conditions
    • Pharmacology: artificial insulin production, drug delivery to
    target sites
    • Medicine: gene therapy, antiviral therapy, vaccination,
    synthesizing clotting factors
    • Other uses: fluorescent fishes, glowing plants etc

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