Wikiomics:Site Directed Mutagenesis

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Revision as of 00:01, 20 November 2007 by Bill Flanagan (Talk | contribs)
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This page is beta. please feel free to update it.

The idea here is to create a flow of work to simulate the point mutation of a single amino acid in a protein.

  1. First step is to choose your target. (This is up to you).
  2. Finding the nucleic sequence (ENTREZ)
  3. What type of mutation will be introduced ? (substitution / deletion / addition)
  4. Can we find the wanted mutation in other related proteins ? (Blast)
  5. How conserved is the protein we wish to mutate ?
  6. Does the w.t. protein has a known 3D structure ? (Check PDB)
    1. if not check out STRUCTURE PREDICTION FLOWCHART
  7. What are the known domains inside our protein ?
    1. see Searching for 3D functional sites in a protein structure
    2. Where is the Active Site ? (SARIG)
    3. What is the location of the mutation? (surface / inside the protein)
  8. What impact does the mutation have on the protein (no backbone changes yet)
  9. If the protein is a multiple subunit protein - what are the subunit interaction residues ? (CSU)
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