Recombinant DNA Method-Mutagenesis

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Recombinant DNA Method-Mutagenesis

• By Jessica Green

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Mutagenesis

• Mutagenesis (the creation or formation of a
  mutation) can be used as a powerful genetic tool.
  By inducing mutations in specific ways and then
  observing the phenotype of the organism the
  function of genes and even individual nucleotides
  can be determined.

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Mutagenesis Laboratory
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Some types of Mutagenesis

• Directed Mutagenesis
• Site-directed/Site-specific Mutagenesis
• Mismatched Mutagenesis

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Directed Mutagenesis

• a largely discredited hypothesis proposing that
  organisms can respond to environmental stresses
  through directing mutations to certain genes or
  areas of the genome.

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Site-directed Mutagenesis

• Where a specific site in a cloned DNA needs to be
  altered in a precise, pre-determined way
• Can be designed to create specific nucleotide
  substitutions, deletions, and so on

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Image of Site-directed Mutagenesis
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Two different forms of Site-specific Mutagenesis

• 5 add-on mutagenesis
• - Diagram on page 148 of the text
• - a new sequence or chemical group is added
  to the 5 end of a PCR product
• Mismatched primer mutagenesis
• - primer is designed to be only partially
  complementary

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Mismatched Mutagenesis

• Diagram pg. 147 of text Figure 5.19
• Can create a desired point mutation at a unique
  predetermined site within a cloned DNA molecule
• At the intended mutation site it bears a base
  that is complementary

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References

• http//humogef.ifj.edu.pl/zaklad/laborator.html
  (pictures)
• Textbook
• http//www3.interscience.wiley.com/cgi-bin/jhome/1
  0009058?CRETRY1SRETRY0