Bradshaw M, Goodnough M C, Johnson E A
Department of Food Microbiology and Toxicology, University of Wisconsin, Madison, Wisconsin, 53706, USA.
Plasmid. 1998 Nov;40(3):233-7. doi: 10.1006/plas.1998.1366.
An RP4-oriT shuttle vector pJIR1457 originally developed for Clostridium perfringens was successfully transferred by conjugation from Escherichia coli to Clostridium botulinum type A strains and to a nontoxigenic C. botulinum type A-transposon Tn916 mutant strain lacking the entire toxin gene cluster. The light chain (LC) of botulinum toxin was highly expressed in the toxin deletion mutant strain from a pJIR1457 construct containing the recombinant botulinal gene for LC. This shuttle vector system will be valuable for genetic analysis of C. botulinum and will enable genetic manipulation and recombinant expression studies of botulinum neurotoxins as pharmaceutical agents.
一种最初为产气荚膜梭菌开发的RP4-oriT穿梭载体pJIR1457，通过接合作用成功地从大肠杆菌转移至A型肉毒梭菌菌株以及一株缺乏完整毒素基因簇的无毒A型肉毒梭菌转座子Tn916突变株。肉毒毒素轻链（LC）在来自含有重组肉毒杆菌轻链基因的pJIR1457构建体的毒素缺失突变株中高表达。该穿梭载体系统对于肉毒梭菌的遗传分析将具有重要价值，并将使肉毒神经毒素作为药物制剂的遗传操作和重组表达研究成为可能。
