Induction of Temperature Sensitive Lambda Repressor
Transform your plasmid DNA into N4830-1 cells (these cells have the gene for the temp. sensitive repressor CI857)
NOTE: Your protein should be cloned into a vector which has the lambda promoter- the PRIT vectors have this.
Transform also a control plasmid that has the lambda promoter and the fusion carrier but not a fusion partener. Incubate at 30ºC (non-permissive) overnight.
Pick one colony into 2mL LB AMP and grow to saturation O/N at 30ºC.
Shift temperature to 42ºC (permissive temp) by mixing with an equal volume of LB AMP pre-heated to 54ºC.
Incubate at 42ºC for 80 minutes.