摘要
目的探索生物膜形成能力、mazEF3,6,9系统的表达与北京基因型结核分枝杆菌生存力的相关性。方法检测北京、非北京基因型菌株与标准毒力株H37Rv在不同培养条件下的生存力、生物膜形成能力。采用PCR技术扩增上述菌株的mazE3,6,9系统,并进行基因测序;使用荧光定量PCR检测所有菌株的mazEF3,6,9基因、mazF3,6,9毒素基因和mazEF3,6,9抗毒素基因的mRNA表达水平;利用WesternBlot技术检测MazF9蛋白表达水平。结果北京基因型菌株在不同培养条件下的生存力、生物膜形成能力高于非北京基因型菌株。北京、非北京基因型菌株mazF3基因193bp处发生碱基突变,由C突变为T。在mRNA表达水平上,与H37Rv相比,北京、非北京基因型菌株中mazE3低表达;非北京菌株中mazE6低表达,mazE9高表达(P<0.05)。与非北京基因型菌株相比,北京基因型菌株中mazEF3高表达、mazE9低表达、mazF3、mazF6、mazF9高表达,mazF9蛋白高表达(P<0.05)。结论北京基因型菌株具有较强的生存力和生物膜形成能力。mazEF系统的差异表达、生物膜成膜能力强可能与北京基因型菌株生存能力强相关。
Objective To explore the correlation of expression of mazEF3, 6, 9 system, biofilm formation ability with the viability of Beijing genotype mycobacterium tuberculosis. Methods Growth profiles and biofilm-formation abilities of Beijing, non-Beijing genotype strains and standard virulence strain H37Rv under different culture conditions were monitored. MazEF3, 6, 9 systems of the above strains were amplified by polymerase chain reaction (PCR), and the genes were sequenced. And then, the mRNA expression of mazEF3, 6, 9,the mazF3, 6 and 9 toxin genes, the mazE3, 6 and 9 antitoxin genes in all strains were detected using Quantitative real time-PCR(qRT-PCR) and expression of the MazF9 was determined by Western Blot. Results The results showed that the viability and biofilm formation ability of Beijing genotype strains were higher than that of non-Beijing genotype strains in different culture conditions. MazF3 gene of Beijing and non-Beijing genotype strains had a base mutation, changed from C to T at 193 bp. Compared with H37Rv, Beijing and non-Beijing genotype strains showed lower expression of mazE3;non-Beijing genotype strains showed lower expression of mazE6 ,high expression of mazE9(P<0.05). Beijing genotype strains also showed higher expression of mazEF3 gene, lower expression of mazE9 and higher expression of mazF3, mazF6 and mazF9 in mRNA expression levels, higher expression of mazF9 at protein level when compared to non-Beijing genotype strains(P<0.05). Conclusion It can be concluded that Beijing genotype strains have strong viability and biofilm forming ability. MazEF system differential expression and strong biofilm-formation abilities may be related to the viability of Beijing genotype strains.
作者
谢婉莹
赵继利
刘微
杨赛丽
袁俐
Xie Wanying;Zhao Jili;Liu Wei;Yang Saili;Yuan Li(Department of Pathogenic Biology and Immunology,School of Medicine,Shihezi University,Shihezi,Xinjiang 832000,China)
出处
《石河子大学学报（自然科学版）》
CAS
北大核心
2018年第6期661-668,共8页
Journal of Shihezi University(Natural Science)
基金
国家自然科学基金项目(81560264)
