Trypsinogen shuffling.

Constructs were made from a first set of trypsinogen modules (mod1, the junction between modules 2 to 3 is agtg) or a second set of modules (mod2, junction between modules 2 to 3 is agtc). White and blue are the total number of colonies obtained per transformation (extrapolated from the number of colonies obtained per plate). All restriction-ligations were performed using equimolar amount of insert and vector except for ts1 that was made using three times less vector (50 ng) than insert. Dimers were identified by running uncut DNA on an agarose gel. Programs used either 6 hours at 37°C (37°C 6 hr) or 50 cycles (conditions given in program); all programs are followed by digestion 5 min at 50°C and heat inactivation 5 min at 80°C. hc, use of high concentration ligase.