广西医科大学学报2019，Vol.36Issue(8)：1215-1221,7.DOI:10.16190/j.cnki.45-1211/r.2019.08.001
鸟分枝杆菌耐受表面活性物质基因的筛选及其相关功能分析
Screening and characterization of Mycobacterium avium gene related to surfactant stress tolerance
摘要
Abstract
Objective:To understand the killing (inhibiting) mechanism of Mycobacterium avium (MAV) adapting to lung environment by screening of genes which resistant to surfactant stress in MAV.Methods:The MAV genomic library was constructed and the sodium dodecyl sulfate (SDS)-tolerant clones were screened from the genomic library by bioactivity level screening strategy,then the complete ORF of the clone and the secondary structure characteristics of its coding protein were analyzed by bioinformatics.The ORF sequence was linked with the pGEX-4T-3 plasmid and transformed into E.coli to construct a recombinant strain which enabled to overexpress the target protein.The survival rates of recombinant strain were detected under the conditions of SDS,H2 O2,NaNO2 and nitroso glutathione (GSNO) respectively.Results:A 1.18 × 104CFU/mL genomic library of MAV was successfully constructed.A SDS-tolerant gene named MAV-4012 was screened from the library.Bioinformatics analysis showed that MAV-4012 protein was a transmembrane protein without signal peptide and its secondary structure was mainly alpha-helix.The survival rate of recombinant strain which overexpressed MAV-4012 in vitro was significantly higher than that of the control strain under 2％ SDS,5 mmol/L H2O2,20 mmol/L NaNO2and GSNO conditions (P ＜ 0.05).Conclusion:MAV-4012 is a novel gene from MAV which resistants to surfactant stress.The protein encoded by MAV-4012 is involved in the stress response of pulmonary surfactant,reactive oxygen and reactive nitrogen,and is an important virulence factor of MAV.关键词
鸟分枝杆菌/MAV-4012/表面活性物质/毒力因子Key words
Mycobacterium avium/MAV-4012/surface-active substance/virulence factor分类
医药卫生引用本文复制引用
宁雪萍,田爽,张依,梁家毓,凌敏..鸟分枝杆菌耐受表面活性物质基因的筛选及其相关功能分析[J].广西医科大学学报,2019,36(8):1215-1221,7.基金项目
The present study was supported by the National Natural Science Foundation of China (No.81260245) and Guangxi Natural Science Foundation (No.2015GXNSFAA139077). （No.81260245）
