pgl gene knock-in

The pgl gene was successfully integrated into the chromosome of the pgl deficient E. coli expression strain (E. coli T7E1, please see above). The corresponding PGL enzyme catalyzes the hydrolysis of 6-phosphogluconolactone to 6-phosphogluconate and thus prevents the accumulation of the electrophile substrate. A loss of the enzyme activity will result in random gluconoylation of cellular proteins including recombinant ones. The optimized strain T7E2 shows no gluconoylation of heterologous expressed protein at all, while in contrast the wildtype shows 6.4%undesired gluconoylation.

Voir les Kits : Red/ET recombination