摘要
目的研究原核类泛素（Pup）-蛋白酶体系统对分枝杆菌生长活力的影响。方法分别敲除耻垢分枝杆菌Pup-蛋白酶体系统中pup和蛋白酶体β亚基基因（prcB）。选取待敲除基因的上下游侧翼序列设计引物，构建自杀性载体，靶基因同源序列和菌株基因组发生同源重组后获得基因敲除的菌株。分别连续测定野生型和基因敲除株在正常及低氧和厌氧应激培养条件下的吸光度值，比较菌株生长活力的变化。结果正确构建敲除pup和prcB基因的突变株，分别命名为△SM-Pup和△SM-prcB。△SM-Pup菌株生长速度快于野生型菌株和△SM—preB株。prcB基因的敲除对细菌的生长活力没有产生明显的影响。结论Pup-蛋白酶体系统对耻垢分枝杆菌的生长产生了一定作用，但具体机制仍需要进一步研究探讨。
Objective To study the effects of prokaryotic ubiquitin-like protein （Pup） -proteasome system on the growth of Mycobacterium strains. Methods The genes encoding Pup （pup gene） and proteasome 13 subunit （prcB gene） were respectively knocked out from Mycobacterium smegmatis （M. sin） strains by homologous recombination. The growth and viability of the wild-type and mutant strains of M. sm were analyzed under normal culture condition and under hypoxia as well as anaerobic conditions. Results The pup and prcB genes were completely and precisely knocked out from M. sm strains and the mutant strains were named △ SM-Pup and △ SM-prcB, respectively. The △ SM-Pup strains grew faster than the wild type （WT） and △SM-prcB strains. No significantly differences in the growth of M. sm were found between the WT and △SM-preB strains. Conclusion The Pup-proteasome system was involved in the growth of M. sin, especially the pup gene. There was difference between pup and prcB genes in regulating the growth of M. sin. The functions and influences of Pup-proteasome system still need further investigation.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2015年第11期832-835,共4页
Chinese Journal of Microbiology and Immunology
基金
国家自然科学基金（81470091）
首都卫生发展科研专项（首发20144-2163）
北京市医院管理局临床医学发展专项经费资助（ZYLX201304）
关键词
分枝杆菌
基因敲除
类泛素
蛋白酶体
生长活力
Mycobacterium
Gene knockout
Prokaryotic ubiquitin-like protein
Proteasome
Viability
