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    Gene Splicing

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    Rhea Joy C. Morales
    Gene splicing is a process where DNA fragments from different organisms are combined or cut and spliced together. In 1973, the first genetically modified organism was created by Herbert Boyer and Stanley Cohen. Gene splicing involves using restriction enzymes to cut DNA at specific sites, then ligases to join the fragments together into a new recombinant DNA molecule, which is then inserted into a host cell. Plasmids are small, circular DNA molecules that are often used as vectors to carry foreign DNA fragments into host cells through processes like transformation, transduction, or conjugation. Nitrosamines are carcinogenic compounds that can form when amines in foods react with nitrites or nitrates added as preservatives, posing health risks if proper

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    Gene Splicing

    Uploaded by

    Rhea Joy C. Morales
    86 views3 pages
    Gene splicing is a process where DNA fragments from different organisms are combined or cut and spliced together. In 1973, the first genetically modified organism was created by Herbert Boyer and Stanley Cohen. Gene splicing involves using restriction enzymes to cut DNA at specific sites, then ligases to join the fragments together into a new recombinant DNA molecule, which is then inserted into a host cell. Plasmids are small, circular DNA molecules that are often used as vectors to carry foreign DNA fragments into host cells through processes like transformation, transduction, or conjugation. Nitrosamines are carcinogenic compounds that can form when amines in foods react with nitrites or nitrates added as preservatives, posing health risks if proper

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    Gene splicing is a process where DNA fragments from different organisms are combined or cut and spliced together. In 1973, the first genetically modified organism was created by Herbert Boyer and Stanley Cohen. Gene splicing involves using restriction enzymes to cut DNA at specific sites, then ligases to join the fragments together into a new recombinant DNA molecule, which is then inserted into a host cell. Plasmids are small, circular DNA molecules that are often used as vectors to carry foreign DNA fragments into host cells through processes like transformation, transduction, or conjugation. Nitrosamines are carcinogenic compounds that can form when amines in foods react with nitrites or nitrates added as preservatives, posing health risks if proper

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    © All Rights Reserved
    Download as DOCX, PDF, TXT or read online from Scribd
    Download as docx, pdf, or txt
    86 views3 pages

    Gene Splicing

    Uploaded by

    Rhea Joy C. Morales
    Gene splicing is a process where DNA fragments from different organisms are combined or cut and spliced together. In 1973, the first genetically modified organism was created by Herbert Boyer and Stanley Cohen. Gene splicing involves using restriction enzymes to cut DNA at specific sites, then ligases to join the fragments together into a new recombinant DNA molecule, which is then inserted into a host cell. Plasmids are small, circular DNA molecules that are often used as vectors to carry foreign DNA fragments into host cells through processes like transformation, transduction, or conjugation. Nitrosamines are carcinogenic compounds that can form when amines in foods react with nitrites or nitrates added as preservatives, posing health risks if proper

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    © All Rights Reserved
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    1.

    GENE SPLICING
    Is a process in which fragments of DNA from one or more organisms are combined; or
    cutting the DNA of a gene to add base pairs.
    It began getting world wide attention in 1973, when the first genetically modified organism
    was recreated.
    Herbert W. Boyer and Stanley N. Cohen created the first DNA recombinant organism.
    Paul Berg created the first DNA recombinant molecule.

    Important Details
    Genetic engineering involves the manipulations of genes in organisms.
    Normally happens prior to the mRNA translation
    The virus or plasmid vectors carry the DNA fragments into cells.
    If genetic material from other species is added to the host, it is called transgenic.
    If genetic material from the same species or species that can naturally breed with the host
    is added, it is called cisgenic.
    Scientist are able to cut and attached fragments of DNA by using restriction enzymes.
    Fragments are attached by using ligase.
    DNA is usually cut at the recognition points on the DNA strand.

    Gene Splicing Mechanism


    There are serval types of common alternative splicing events:
    Exon Skipping
    - This is the most common known alternative splicing mechanism in which
    exon(s) are included or excluded from the final gene transcript leading to
    extended or shortened mRNA variants.
    - Its purpose is to restore the reading frame within the gene.
    - Mainly used to help with Duchenne muscular dystrophy.

    Intron Retention
    - An event in which an intron is retained in the final transcript. In humans 2-5 %
    of the genes have been reported to retain introns.
    Alternative 3' splice site and 5' splice site
    - Alternative gene splicing includes joining of different 5' and 3' splice site. In
    this splicing mechanism, two or more alternative 5' splice site compete for
    joining to two or more alternate 3' splice site.

    Gene Splicing Process


    Step 1: Plasmid is extracted from the bacteria, and the gene of interest is targeted.
    Step 2: The gene of interest is identified and isolated.
    Step 3: Restriction enzymes are added to target the specific section in the gene sequence so
    that they are separated from rest of the strand.
    Step 4: The gene is then linked together with the plasmid by using ligase.
    Step 5: The recombinant DNA is then re-inserted in the bacteria and then the bacteria is
    inserted into the host organism.

    2. Plasmid
    A circular form of DNA often used as a vector in genetic engineering.
    A small DNA molecule within a cell that is physically separated from a chromosomal DNA
    and can replicate independently. They are most commonly found in bacteria as small circular,
    double-stranded DNA molecules; however, plasmids are sometimes present in archaea and
    eukaryotic organisms.
    Are considered replicons, a unit of DNA capable of replicating autonomously within a suitable
    host. However, plasmids, like viruses, are not generally classified as life. Plasmids can be
    transmitted from one bacterium to another (even of another species) via three main
    mechanisms: transformation, transduction, and conjugation. This host-to-host transfer of
    genetic material is called horizontal gene transfer, and plasmids can be considered part of the
    mobilome.

    Plasmid three main feature


    - Multiple cloning site
    - Origin of replication
    - Selectable marker

    3. Nitrosamines
    Nitrosamines are important group of mutagens found in tobacco, and may also be formed in
    smoked meats and fish via the interaction of amines in food with nitrites added as preservatives.
    Sleiman identified the formation of carcinogenic tobacco-specific nitrosamines (TSNAs) from
    the reaction of adsorbed nicotine with nitrous acid.
    Nitrosamines are formed by reaction of secondary or tertiary amines with a nitrosating agent.
    In foods, the nitrosating agent is usually nitrous anhydride, formed from nitrite in acidic,
    aqueous solution. Food constituents and the physical make-up of the food can effect
    nitrosamine formation. Ascorbic acid and sulfur dioxide are used to inhibit nitrosamine
    formation in foods.
    The possible formation of nitrosamines from amine present in or derived from the diet occurs
    by reaction with nitrous acid at pH 4. In man, gastric juice attains a pH of 1.1. Such high
    concentrations of hydrogen ions give rise to nitrosyl cation NO , a highly reactive nitrosating
    agent. The presence of meat, myoglobin, or hemoglobin serve to neutralize this cation by
    reacting with Fe 2 . Fish and cheese do not provide this neutralization, hence permitting the
    nitrosation of the present amines to possible carcinogenic nitrosamines.

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