Wikiomics:Site Directed Mutagenesis

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 Wikiomics: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)