Secretory expression of recombinant diphtheria toxin mutants in B. Subtilis.
Three diphtheria toxin (DT) mutants CRM-197, DT-del (148) and DT-E148S-K516A-F530A were cloned in B. Subtilis plasmid PSM604 under the subtilisin signal sequence. The expression was effective in both SMS300 and SMS118, but higher yield of 7.1 mg/L was observed in SMS300 compared with 2.1 mg/L in SMS118. Western blot showed that the recombinant protein could be effectively secreted into the culture medium as a 58 ku peptide, and could be degraded into two peptides of 37 ku and 21 ku.
Zhou
J
J
Department of Pediatrics, Tongji Hospital, Tongji Medical University, Wuhan 430030.
Petracca
R
R
D001412:Bacillus subtilis / Q000235:genetics; D001426:Bacterial Proteins; D004167:Diphtheria Toxin / Q000096:biosynthesis; D010957:Plasmids; D011994:Recombinant Proteins / Q000235:genetics
D001426:Bacterial Proteins; D004167:Diphtheria Toxin; D011994:Recombinant Proteins; C028581:CRM197 (non-toxic variant of diphtheria toxin)