GenBio2SumTest1

    Cards (21)

    • Genetic engineering
      a gene modification process wherein the Deoxyribonucleic acid (DNA) is transferred from one organism to another.
    • Classical breeding
      Focuses on the mating of organisms with desirable qualities or traits
    • Genetic engineering
      Involves molecular techniques to modify the traits of a target organism.
    • Genetically modified organisms
      Have been subject for public scrutiny whether it is safe to use or ethically accepted.
    • 4 Processes of Genetic Engineering
      DNA Cleavage, Production of Recombinant DNA, Cloning, Screening
    • Stage 1. DNA Cleavage
      A restriction endonuclease is used to cleave the source DNA into a different set of fragments. The endonuclease’s recognition sequence is likely to occur many times within the source of DNA, thus cleavage will produce a large number of different fragments. The fragments can be separated from one another according to their size by gel electrophoresis.
    • Stage 2. Production of Recombinant DNA
      The fragments of DNA are inserted into plasmids or viral vectors that have been cleaved with the same restriction endonuclease as the source DNA.
    • Stage 3. Cloning
      The plasmids or viruses serve as vectors that can introduce the DNA fragments into cells--- usually, but not always bacteria. As each cell produces, it forms a clone of cells that all contain the fragment-bearing vector.
    • Stage 4. Screening
      The clones containing a specific DNA fragment of interest are identified from the clone library
    • Methods of Introducing Plasmids into the Host Organism
      Biolistics, Plasmid insertion by Heat Shock Treatment, Electroporation
    • Methods to Screen Recombinant Cells
      Selection of plasmid DNA containing cells, Selection of transformed cells with the desired gene, Polymerase Chain Reaction (PCR) detection of plasmid DNA
    • PCR reactions specific for plasmid sequences will confirm/identify the type of plasmid used for the transformation through the following steps:
      Step 1. Denaturation, Step 2. Annealing of Primers, Step 3. Elongation
    • Biolistics
      This technique uses a “gene gun” to fire DNA-coated pellets on plant tissues. Cells that are able to survive and take up the expression plasmid coated pellets can acquire the ability to express the designed protein.
    • Plasmid insertion by Heat Shock Treatment
      is a process used to transfer plasmid DNA into bacteria
    • Electroporation
      This technique follows a similar methodology as Heat Shock Treatment but uses electric shock to expand the membrane pores. This method is commonly used for the insertion of genes into mammalian cells.
    • Selection of plasmid DNA containing cells
      A selection marker within the inserted plasmid DNA sequence allows the selection of “transformants”.
    • Selection of transformed cells with the desired gene
      The most general procedure for screening clone libraries to find a particular gene is hybridization - the cloned genes form base-pairs with complementary sequences on another nucleic acid.
    • Polymerase Chain Reaction (PCR) detection of plasmid DNA

      the presence of the desired gene in the inserted plasmids may also be confirmed through PCR amplification
    • Step 1. Denaturation
      An excess primer, synthetic sequence of 20 to 30 nucleotides
    • Step 2. Annealing of Primers
      The solution is allowed to cool to about 60°C so that the single strands of DNA reassociate into double strands.
    • Step 3. Elongation
      Using the primer, the polymerase copies the rest of the fragment and both DNA strands are replicated resulting into two copies of the original fragment
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