Wikiomics:Site Directed Mutagenesis
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| - | This page is beta. | + | 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. | The idea here is to create a flow of work to simulate the point mutation of a single amino acid in a protein. | ||
| - | #First step is to choose your target. (This is up to you | + | #First step is to choose your target. (This is up to you.) |
#Finding the nucleic sequence ([http://www.ncbi.nlm.nih.gov/entrez/query.fcgi ENTREZ]) | #Finding the nucleic sequence ([http://www.ncbi.nlm.nih.gov/entrez/query.fcgi ENTREZ]) | ||
#What type of mutation will be introduced ? (substitution / deletion / addition) | #What type of mutation will be introduced ? (substitution / deletion / addition) | ||
#Can we find the wanted mutation in other related proteins ? (Blast) | #Can we find the wanted mutation in other related proteins ? (Blast) | ||
#How conserved is the protein we wish to mutate ? | #How conserved is the protein we wish to mutate ? | ||
| - | #Does the w.t. protein | + | #Does the w.t. protein have a known 3D structure ? (Check [http://www.rcsb.org/pdb/ PDB]) |
| - | ## | + | ##If not, check out [http://speedy.embl-heidelberg.de/gtsp/flowchart2.html STRUCTURE PREDICTION FLOWCHART] |
#What are the known domains inside our protein ? | #What are the known domains inside our protein ? | ||
## see [[Wikiomics:Searching for 3D functional sites in a protein structure]] | ## see [[Wikiomics:Searching for 3D functional sites in a protein structure]] | ||
## Where is the Active Site ? ([http://bioinfo2.weizmann.ac.il/~pietro/SARIG/V3/index.html SARIG]) | ## Where is the Active Site ? ([http://bioinfo2.weizmann.ac.il/~pietro/SARIG/V3/index.html SARIG]) | ||
## What is the location of the mutation? (surface / inside the protein) | ## What is the location of the mutation? (surface / inside the protein) | ||
| - | #What impact does the mutation have on the protein (no backbone changes yet) | + | #What impact does the mutation have on the protein? (no backbone changes yet) |
#If the protein is a multiple subunit protein - what are the subunit interaction residues ? (CSU) | #If the protein is a multiple subunit protein - what are the subunit interaction residues ? (CSU) | ||
[[Category:Protocol]] [[Category:In silico]] [[Category:Cloning]] | [[Category:Protocol]] [[Category:In silico]] [[Category:Cloning]] | ||
Current revision
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.
- First step is to choose your target. (This is up to you.)
- Finding the nucleic sequence (ENTREZ)
- What type of mutation will be introduced ? (substitution / deletion / addition)
- Can we find the wanted mutation in other related proteins ? (Blast)
- How conserved is the protein we wish to mutate ?
- Does the w.t. protein have a known 3D structure ? (Check PDB)
- If not, check out STRUCTURE PREDICTION FLOWCHART
- What are the known domains inside our protein ?
- see Wikiomics:Searching for 3D functional sites in a protein structure
- Where is the Active Site ? (SARIG)
- What is the location of the mutation? (surface / inside the protein)
- What impact does the mutation have on the protein? (no backbone changes yet)
- If the protein is a multiple subunit protein - what are the subunit interaction residues ? (CSU)
