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    Aspects of Genetic Engineering 2.4

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    Dequan
    The document discusses genetic engineering techniques such as restriction enzymes and DNA ligase that are used to insert genes into plasmids. It describes how restriction enzymes cut DNA at specific sequences and DNA ligase joins DNA molecules. The steps of recombinant DNA technology including isolating genes, restriction enzyme digestion, ligation, and inserting recombinant DNA into a host are outlined. Applications like producing human insulin in bacteria and approaches to gene therapy are also summarized.

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    Aspects of Genetic Engineering 2.4

    Uploaded by

    Dequan
    15 views18 pages
    The document discusses genetic engineering techniques such as restriction enzymes and DNA ligase that are used to insert genes into plasmids. It describes how restriction enzymes cut DNA at specific sequences and DNA ligase joins DNA molecules. The steps of recombinant DNA technology including isolating genes, restriction enzyme digestion, ligation, and inserting recombinant DNA into a host are outlined. Applications like producing human insulin in bacteria and approaches to gene therapy are also summarized.

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    The document discusses genetic engineering techniques such as restriction enzymes and DNA ligase that are used to insert genes into plasmids. It describes how restriction enzymes cut DNA at specific sequences and DNA ligase joins DNA molecules. The steps of recombinant DNA technology including isolating genes, restriction enzyme digestion, ligation, and inserting recombinant DNA into a host are outlined. Applications like producing human insulin in bacteria and approaches to gene therapy are also summarized.

    Copyright:

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

    Aspects of Genetic Engineering 2.4

    Uploaded by

    Dequan
    The document discusses genetic engineering techniques such as restriction enzymes and DNA ligase that are used to insert genes into plasmids. It describes how restriction enzymes cut DNA at specific sequences and DNA ligase joins DNA molecules. The steps of recombinant DNA technology including isolating genes, restriction enzyme digestion, ligation, and inserting recombinant DNA into a host are outlined. Applications like producing human insulin in bacteria and approaches to gene therapy are also summarized.

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    © All Rights Reserved
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    Genetics,

    Variation and
    Natural Selection
    Genetic Engineering

    Done by: Dequan Holder


    Restriction Enzymes and DNA Ligase

    • Restriction enzymes are DNA-cutting enzymes. Each enzyme recognizes


    one or a few target sequences and cuts DNA at or near those sequences.
    • Many restriction enzymes make staggered cuts, producing ends with
    single-stranded DNA overhangs. However, some produce blunt ends.
    • DNA ligase is a DNA-joining enzyme. If two pieces of DNA have matching
    ends, ligase can link them to form a single, unbroken molecule of DNA.
    • In DNA cloning, restriction enzymes and DNA ligase are used to insert
    genes and other pieces of DNA into plasmids.
    • Restriction enzymes and DNA ligase are often used to insert genes and
    other pieces of DNA into plasmids during DNA cloning.
    Diagram depicting restriction process.
    Diagram depicting the DNA ligase process.
    Steps in recombinant DNA (rDNA) technology.

    • Recombinant DNA (rDNA) technology refers to the process of joining


    DNA molecules from two different sources and inserting them into a
    host organism, to generate products for human use. This process
    consist of a number of steps which are:
    • Isolation of genes
    • Restriction enzyme digestion
    • Cloning of genes
    • Ligation of DNA
    • Insertion of recombinant Dna into the host
    Isolation of Genes

    • Gene isolation, which is the isolation of a particular gene from the


    rest for use in genetic engineering.
    • The cell containing the gene is opened and the DNA is purified.The
    gene is separated by using restriction enzymes to cut the DNA into
    fragments or polymerase chain reaction (PCR) to amplify up the
    gene segment.
    Restriction Enzyme Digestion

    • Restriction enzymes act as molecular scissors that cut DNA at


    specific locations.
    • They involve the incubation of the purified DNA with the selected
    restriction enzyme, at conditions optimal for that specific enzyme.
    Ligation of DNA Molecules

    • The purified DNA and the vector of interest are cut with the same
    restriction enzyme.
    • This gives us the cut fragment of DNA and the cut vector, that is
    now open.
    • The process of joining these two pieces together using the enzyme
    ‘DNA ligase’ is ‘ligation’. The resulting DNA is ‘recombinant DNA‘.
    Insertion of Recombinant DNA Into Host

    • The recombinant DNA is introduced into a recipient host cell. This


    process is ‘Transformation’.
    • Bacterial cells do not accept foreign DNA easily. Therefore, they
    are treated to make them ‘competent’ to accept new DNA
    An Application of Genetic Engineering

    Insulin Production
    Bacterial cells can be genetically modified so that they have the
    gene for producing human insulin. As these modified bacteria grow,
    they produce human insulin. This protein can be purified and
    supplied to diabetics.
    Gene Therapy
    Gene therapy is an experimental technique that uses genes to treat or
    prevent disease. There are three (3) known approaches to gene therapy
    which are:
    • Replacing a mutated gene that causes disease with a healthy copy of the
    gene.
    • Inactivating, or “knocking out,” a mutated gene that is functioning
    improperly.
    • Introducing a new gene into the body to help fight a disease.
    Although gene therapy is a promising treatment option for a number of
    diseases (including inherited disorders, some types of cancer, and certain
    viral infections), the technique remains risky and is still under study.
    Application in Gene Therapy.

    • Cystic fibrosis results from a mutation in the carrier protein (CFTR)


    which pumps chloride ions out of cells in the respiratory tract and
    alimentary canal.
    • Person who are homozygous recessive for the disorder have thick
    and sticky mucus.
    • Liposomes and viruses have been used as vectors to deliver the
    dominant allele into the cells of respiratory treact using an inhaler.
    • This had mixed success as the vectors tend to stimulate
    immflammation and some clinical trials had to be stopped.
    What are GMOs?
    • GMO stands for “Genetically Modified Organism,” and the phrase refers
    to plants or animals that have been genetically engineered to produce a
    specific set of characteristics.
    • DNA is taken from a completely different species (like a virus or fish) and
    inserted into a crop (like a carrot) to transfer genetic traits. These
    experimental combinations of genes do not occur in nature.
    • The use of GMOs in the environment also comes with implications which
    can be medical, agricultural, ethical and social.
    Implications involving the usage of GMOs in
    the Environment.

    Agriculturally
    • Out-crossing is the unintentional breeding of a domestic crop with a
    related plant.
    • A major environmental concern associated with GM crops is their
    potential to create new weeds through out-crossing with wild relatives.
    • Managing weeds is one of the most tedious tasks of farming.
    • scientists developed genetically modified herbicide-tolerant (HT) crops
    so farmers can spray their fields with weed killers without affecting their
    crop yield.This led to the use of glyphosate herbicides. Overtime the
    weeds developed aresistance to the herbicide.  
    Implications involving the usage of GMOs in
    the Environment cont’d

    Ethically
    • Antibiotic resistance factors used to identify GMOs could escape
    and be transferred to pathogenic organisms.
    • Herbicide resistance genes could be transferred from pollen to
    weed species and lead to the development of superweeds that are
    resistant to herbicides.
    • Foreign genes can pollute non-GM and organic crops which require
    certification that they provide GM-free food.
    Implications involving the usage of GMOs in
    the Environment cont’d

    Medically
    • For many years diabetes was treated by regular injections of
    insulin which was extracted from animals.but now it is produced
    by GM bacteria or yeasts.
    • The main disadvantage of GM human insulin is that some people
    • Have reported that they do not experience any warning signs of a
    hypoglycaemic attack when the blood glucose concentration falls
    increasing the likelihood of a diabetic coma.
    END OF PRESENTATION!!!

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