Talk:20.109(S13):Design mutant (Day1)

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Revision as of 12:34, 10 March 2013

20.109(S13): Laboratory Fundamentals of Biological Engineering

Home        Schedule Spring 2013        Assignments       
DNA Engineering        Protein Engineering        Cell Engineering              

Mutant choices for positive control

The mutants below were chosen based on promising student results in previous years, then tested by the teaching faculty.

Mutation X#Z Primer (forward) Silent mutation New restriction site Known effect
D24H C GAC AAG GAT GGG CAC GGT ACC ATC ACC -ANS Gly25 KpnI Affinity decrease and cooperativity increase (both substantial)
E67K GGC AAT GGT ACG ATT TAC TTT CCT AAA TTT CTT ACT ATG ATG GC Gly63 RsaI Small affinity increase
T79P GA AAA ATG AAG GAC CCA GAC AGC GAA GAG GAA ATC C Thr79 PpuMI Small affinity decrease and perhaps slight cooperativity increase
M124S GAT GAA GAA GTT GAT GAA AGT ATA CGG GAA GCA GAT ATC GAT GG Arg126 AccI Affinity decrease and cooperativity increase


Note that we also investigated D22H and D129P, but both of these turned out to destroy calcium response, and in low salt conditions to eliminate fluorescence itself. The limited choices above aren't meant to imply that you must narrowly investigate these residues or ones just like them.

T/R

==W/F==
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