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    Application of Biotechnology On Genetic Engineering (DNA Cloning)

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    Jerome Magno
    DNA cloning involves making multiple copies of a piece of DNA. It begins by separating the two strands of DNA using enzymes like helicase. Restriction enzymes then cut the DNA at specific sequences. The cut DNA is joined to a vector using ligase enzymes and introduced into host cells. The host cells copy both their own DNA and the vector DNA, producing multiple copies of the inserted DNA fragment. The cloned DNA is then isolated from the host cells. DNA cloning is used to study gene function and characteristics and produce large amounts of proteins.

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    Application of Biotechnology On Genetic Engineering (DNA Cloning)

    Uploaded by

    Jerome Magno
    70 views4 pages
    DNA cloning involves making multiple copies of a piece of DNA. It begins by separating the two strands of DNA using enzymes like helicase. Restriction enzymes then cut the DNA at specific sequences. The cut DNA is joined to a vector using ligase enzymes and introduced into host cells. The host cells copy both their own DNA and the vector DNA, producing multiple copies of the inserted DNA fragment. The cloned DNA is then isolated from the host cells. DNA cloning is used to study gene function and characteristics and produce large amounts of proteins.

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    DNA Cloning

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    DNA cloning involves making multiple copies of a piece of DNA. It begins by separating the two strands of DNA using enzymes like helicase. Restriction enzymes then cut the DNA at specific sequences. The cut DNA is joined to a vector using ligase enzymes and introduced into host cells. The host cells copy both their own DNA and the vector DNA, producing multiple copies of the inserted DNA fragment. The cloned DNA is then isolated from the host cells. DNA cloning is used to study gene function and characteristics and produce large amounts of proteins.

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    70 views4 pages

    Application of Biotechnology On Genetic Engineering (DNA Cloning)

    Uploaded by

    Jerome Magno
    DNA cloning involves making multiple copies of a piece of DNA. It begins by separating the two strands of DNA using enzymes like helicase. Restriction enzymes then cut the DNA at specific sequences. The cut DNA is joined to a vector using ligase enzymes and introduced into host cells. The host cells copy both their own DNA and the vector DNA, producing multiple copies of the inserted DNA fragment. The cloned DNA is then isolated from the host cells. DNA cloning is used to study gene function and characteristics and produce large amounts of proteins.

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    © All Rights Reserved
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    of 4

    St.

    Louis University
    Baguio City, Philippines
    A.Y. 2014-2015

    Application of Biotechnology on
    Genetic Engineering (DNA Cloning)

    Submitted by: Jerome T. Magno


    Submitted to: Engr. Katelyn G. Gabon
    DNA Cloning

    DNA cloning is the starting point for many genetic engineering


    approaches to biotechnology research.

    Large amounts of DNA are needed for genetic engineering.


    Multiple copies of a piece of DNA can be made either by using polymerase
    chain reaction (PCR) or by cloning DNA in cells.

    The polymerase chain reaction (PCR) is a technique that is


    used to make multiple copies of a piece of DNA.PCR is carried out in
    vitro, but mimics what happens in cells when DNA is copied (replicated)
    prior to cell division.

    How does PCR work?

    Before PCR can occur, the two strands in the DNA double helix
    need to be separated. This is called denaturation. It is done by raising
    the temperature of the DNA solution. This causes the hydrogen bonds
    between the complementary DNA chains to break, and the two strands
    separate.

    Next, the temperature is lowered and an enzyme (usually Taq


    polymerase) joins free DNA nucleotides together. The order in which these
    nucleotides are joined to the new strand is determined by the sequence
    of nucleotides in the original DNA strand which is being copied.

    The result is a double stranded DNA molecule which contains one newly
    made strand and one original strand.

    Next, the newly created double helix is separated (by heating the
    solution) and the cycle is repeated.

    How is DNA cloned in cells?

    To get multiple copies of a gene or other piece of DNA you must isolate,
    or cut, the DNA from its source and then paste it into a DNA vector
    that can replicate (or copy) itself.

    The four main steps in DNA cloning are:

    1. The chosen piece of DNA is cut from the source organism using
    restriction enzymes.

    In the laboratory, restriction enzymes (or restriction endonucleases)


    are used to cut DNA into smaller fragments. The cuts are always made at
    specific nucleotide sequences. Different restriction enzymes recognize
    and cut different DNA sequences.
    Like all enzymes, a restriction enzyme works by shape-to-shape matching.
    When it comes into contact with a DNA sequence with a shape that matches
    a part of the enzyme, called the recognition site, it wraps around the
    DNA and causes a break in both strands of the DNA molecule.

    Each restriction enzyme recognizes a different and specific recognition


    site, or DNA sequence. Recognition sites are usually only short - 4-8
    nucleotides.

    2. The piece of DNA is pasted into a vector and the ends of the
    DNA are joined with the vector DNA by ligation.

    In cells and in the lab, enzymes called ligases are used to join fragments
    of DNA together. Only DNA fragments that have matching, complementary
    ends can be joined by ligation.

    Ligases join fragments of DNA together by catalyzing the formation of


    bonds between neighboring nucleotides.

    Ligation in cells

    Cells naturally carry out ligation during DNA replication, when the
    Okazaki fragments are joined together. Cells also use ligation to repair
    DNA that has been damaged, either by normal cell metabolism or by
    environmental factors, such as UV light or radiation. Up to 1 million
    breaks can occur in the DNA of a single human cell each day.

    3. The vector is introduced into a host cell, often a bacterium or


    yeast, by a process called transformation. The host cells copy the
    vector DNA along with their own DNA, creating multiple copies of
    the inserted DNA.

    Bacteria are commonly used as host cells for making copies of DNA in the
    lab because they are easy to grow in large numbers. Their cellular
    machinery naturally carries out DNA replication and protein synthesis.

    4. The vector DNA is isolated (or separated) from the host cells DNA
    and purified.

    DNA that has been cut and pasted from an organism into a vector is
    called recombinant DNA. Because of this, DNA cloning is also called
    recombinant DNA technology.

    What is cloned DNA used for?

    DNA cloning is used to create a large number of copies of a gene or


    other piece of DNA. The cloned DNA can be used to:
    Work out the function of the gene
    Investigate a genes characteristics (size, expression, tissue
    distribution)
    Look at how mutations may affect a genes function
    Make large concentrations of the protein coded for by the gene

    Personal Insights

    Biotechnology has help me to become knowledgeable of the


    things and occurrences in my system and certain processes. One of these
    is the nucleic acids, one of the biological molecules, which have two
    types. One of this is the DNA (Deoxyribonucleic Acid) that we know is
    present in all cells. In, genetic engineering in which it involves DNA
    cloning, my understanding about this topic has broaden and we can call
    it sharp because of my background of Biotechnology. In DNA cloning first
    is we have to separate the two strands in the double helix structure of
    DNA and then we imply an enzyme to join the free nucleotides together
    in order of the original sequence and we learned that in the DNA
    complementary base pairing the adenine (A) is paired to thymine (T),
    Guanine (G) is then paired to Cytosine (C). So in the process of DNA
    cloning the DNA is cut by using restriction enzymes and one example cited
    in our lecture is the enzyme helicase that it separates the 2 DNA strands
    by breaking the weak hydrogen bonds. And after it is introduced into a
    vector and it will imply the process of ligation in which we imply the
    use of an enzyme example is Ligase join fragments of DNA together. And
    we know that in the cell there is a DNA replication, and in this ligation
    carries out when the Okazaki fragments (series of short segments on the
    lagging strand) are joined together. And so it will introduced to a host
    cell or a vector that copy the vector DNA along with their own DNA,
    creating multiple copies of the inserted DNA as well as the DNA
    replication implies. And after the copying the DNA vector is separated
    to the host cells just like the relationship of a substrate and an
    enzyme, the substrate will go to its respective enzyme and when the
    process is completed or one is benefited, the substrate will separate
    into the enzyme and other reaction will the occur. And there is a
    formation of cloned DNA or what we call the recombinant DNA. So, we then
    employed biotechnology terms and applications in understanding the DNA
    cloning. Biotechnology is crucial in our lives, it will open new
    opportunities to understand and develop our world, and it will create
    improvements and innovations to further make our life easier.

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