摘要
菌蜕是革兰氏阴性菌在噬菌体Phix174的溶菌基因E的作用下形成的细菌空壳,通常由λpL/pR-cI857温控系统通过调控E基因的表达制备。通过重叠PCR技术,将λpL/pR-cI857温控系统中λpR启动子的第2个操纵基因(OR2)的第9位碱基由T突变为C(T→C)。溶菌试验结果表明,突变后的λpL/pR-cI857系统在37℃时仍能稳定抑制基因E的表达,对大肠杆菌的溶菌率为99.9%。通过对λpR启动子的改造,扩大了λpL/pR-cI857温控系统的温度调控范围,为菌蜕疫苗的研制提供了参考。
Bacterial ghost is intact envelope of Gram-negative bacteria, which is produced by the function of the lysis gene E from bacteriophage PhiX174. The expression of the lysis gene E is usually controlled by the thermosensitive λpL/pR-cI857 promoter. In this study, we described a mutation （T→C） at the ninth nucleotide of the OR2 in the kpR promoter of theλpL/pR-cI857 system by overlap PCR. The bacteriolytic assay showed that the mutation in the λpL/pR-cI857 system enhanced the temperature of repressing the expression of gene E up to 37 ℃. The lysis efficiency of altered λpR promoter in Escherichia coli DHS5a and avian pathogenic E. coli DE17 was up to 99.9%. The expanded range of temperature will benefit for the production of bacterial ghost.
出处
《生物工程学报》
CAS
CSCD
北大核心
2012年第12期1423-1430,共8页
Chinese Journal of Biotechnology
基金
国家自然科学基金(Nos.31001078
31072161
30871851)资助~~
关键词
大肠杆菌
菌蜕
重叠PCR
突变
Escherichia coli, bacterial ghosts, overlap PCR, mutation
