GTE TRI 1 LECT 5

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  • Created by: Loz1669
  • Created on: 01-01-20 14:31
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  • Genes to ecosystems
    • Recombinant DNA technology
      • Genetic manipulation
        • "manipulating the genotype of organisms to produce a specific phenotype."
      • Applications
        • To gain a DNA sequence information for individual genes of the genomes of organisms. The requirement for this is to make large quantities of DNA - this is often achieved by cloning.
        • To analyse the function of a protein from a particular gene. Required to make large quantities of that specific protein. - this is known as the expression of cloned genes.
        • To analyse the function of individual genes and proteins in cells. Required to get relevant genes into cells or remove genes from cells. These are called transgenic cells
        • To analyse the functions on individual genes in the whole organism. Requirement is to get genes into all and remove genes from whole animals - known as transgenic animals.
          • Analysing the function of the gene
            • Analyse the function of the protein that the gene produces.
              • We can express this gene in bacteria to coerce the quantities of bacteria to make gene products for us
                • We then grow large amounts of bacteria and purify the specific protein
                  • Used in medical production and analysing the function of a protein.
          • Transgenic cell or animals which have had foreign genes introduced into them
            • Ethical issues
            • Human gene therapy
        • To use knowledge of functions of a gene to cure disease. The requirement for this is to understand the roles of genes in tissues of all animals in order to be able to recover the failed function of genes This is known as gene therapy.
      • Cloning
        • The aim of cloning DNA is start with small quantities of DNA and end up with enough to analyse and work with. Usually bacteria is used because it grows fast.
          • Small circles of DNA called plasmids replicate with bacteria and their often hundreds of copies of plasmids in each bacterial cell.
            • These plasmids contain genes for antibiotic resistance which means they can be selected for in order to encourage their presence in culture of bacteria.
              • The mechanism for cloning in human genes is to place that gene within one of these plasmids.
                • These plasmids are known as vectors as they carry foreign DNA
                  • The process of cloning includes cutting and Joining of DNA
                    • Restriction enzymes cut DNA at a specific sequence
                      • DNA can be rejoined using DNA ligase
                    • DNA from the plasmid factor is purified and then cut to linearise the plasmid
                      • DNA is extracted from the organism cell and is cut then joined to the plasmid DNA
                        • This recombinant DNA is formed back to a circle and put back into the bacteria.
                          • This results in a culture of bacteria which each contains a different gene from our organism
                            • This is known as a library of cloned genes
                              • Selecting a specific clone
                                • Genomic libraries contains lots of bacteria each containing a different gene of the DNA insert
                                  • Each bacteria is seperated and then incubated at 37 degrees to produce colonies
                                    • These are then screened to find which contains the gene of interest
                                      • This can then be expanded to the desired quantity
                              • The challenge is to find the correct clone containing the sequence that we want.

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