http://www.nature.com/nrm/journal/v3/n12/pdf/nrm975.pdf
"Protein engineering 20 years on" Brannigan J, Wilkinson A
Combinatorial mutagenesis:
- mix DNA fragments derived from multiple sources (eg. mutants/wild types, or closely related genes) and PCR it up. 'sexual PCR'
- SHIPREC (Sequence Homology Independent Protein Recombination) and ITCHY (Independent Truncation for the Creation of HYbrid enzymes) -> allows distantly related genes to be put together in chimeras. SCRATCHY (shuffling) then uses sexual PCR to further shuffle things up
- Random Insertion Deletion (RID) mutation? Remove arbitrary number of bases and replace with arbitrary number of bases?
http://www.nature.com/nbt/journal/v20/n1/full/nbt0102-76.html
- read "The blind watchmaker" and "The selfish gene" by Richard Dawkins
- phage display?
- application: artificial blood substitutes for blood transfusions (based on re-engineered Haemoglobin)
- long lasting insulin that doesn't clump together and lose its activity
- COMPLETELY NOVEL PROTEINS BASED ON UNNATURAL AMINO ACIDS: DESIGN NEW aminoacyle-tRNA synthetase enzymes to 'incorrectly' activate tRNAs!
http://en.wikipedia.org/wiki/Weasel_program
R Dawkins' Weasel program
Try to arrive at the phrase "Methinks it is like a weasel"
Purely by chance it would take a very long time (26^28 combinations). However, by coupling random changes with a 'fitness advantage' (ie. phrases that more closely resemble MIILAW are selected for), the change is possible
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