目的 揭示结核分枝杆菌易化子超家族(MFS)蛋白Rv0037c的结构和功能特征。方法 应用TM-HMM Server v。2。0、Swiss-model、DN AM AN等对Rv0037c蛋白进行生物信息学分析，在耻垢分枝杆菌中构建PMV261-Rv0037c过表达菌株，获取pMV261/Msm对照菌株，分析其生物学功能。结果 Rv0037c蛋白具有11个跨膜结构域(TMDs)，C/N域由"3+2"螺旋束组成，在分枝杆菌属中非常保守。过表达菌株和对照菌株的生长动力学曲线几乎完全重合，过表达菌株与对照菌株的生长量比较，差异无统计学意义(P＞0。05)。对照菌株的OD600为2。567±0。162，过表达菌株的OD600为2。419±0。456，二者的生物膜生长量比较，差异无统计学意义(P＞0。05)。过表达菌株的溴化乙锭(EB)累积量较对照菌株低，差异有统计学意义(P＜0。001)。两种菌株对各种药物的最低抑菌浓度(MIC)比较，差异无统计学意义(均P＞0。05)。结论 Rv0037c蛋白为分枝杆菌MFS家族蛋白的特殊成员且结构非常保守，Rv0037c过表达对耻垢分枝杆菌生长、菌落形态、滑动能力、生物膜形成和MIC均无影响，但可使EB的累积量显著减少。
Characteristics of structure and functions of Rv0037c protein of Mycobac-terium tuberculosis
Objective To explore the structure and functions of Rv0037c protein,a member of major facilitator su-perfamily(MFS)proteins of Mycobacterium tuberculosis.Methods Bioinformatics analysis on Rv0037c protein was performed with TMHMM Server v.2.0,Swiss-model and DNAMAN software.The PMV261-Rv0037c overex-pressed strain was constructed in Mycobacterium smegmatis,pMV261/Msm control strain was obtained,its biolo-gical function was analyzed.Results Rv0037c protein contained 11 transmembrance domains,the C/N domain con-sisted of"3+2"helical bundles,which was highly conservative in genus Mycobacterium.The growth kinetics curves of the overexpressed strain and the control strain almost completely overlapped,and there was no statistical difference in growth between the overexpressed strain and the control strain(P＞0.05).The OD600 of the control strain was 2.567±0.162,while that of the overexpressed strain was 2.419±0.456,there was no statistical diffe-rence in biofilm growth between the two strains(P＞0.05).The accumulation of ethidium bromide(EB)in overex-pressed strain was lower than that in control strain,difference was statistically significant(P＜0.001).There was no statistically significant difference in the minimum inhibitory concentration(MIC)of two bacterial strains against various drugs(both P＞0.05).Conclusion The Rv0037c protein is a special member of the Mycobacterium MFS family proteins and has a very conserved structure.Overexpression of Rv0037c has no effect on the growth,colony morphology,sliding ability,biofilm formation,and MIC of Mycobacterium smegmatis,but can significantly reduce the accumulation of EB.
万方数据
维普
