【摘要】 结核分枝杆菌引起的结核病危害严重,是传染性较强的全球慢性感染病之一,具有潜伏感染的特性.结核分枝杆菌来源的RNA解旋酶RhlE依赖于ATP水解活性,参与RNA代谢,调节核糖体的装配与降解,但是其催化机制和结构基础尚不清楚.本文对RhlE进行了体外表达纯化、动态光散射和酶活性检测研究.结果表明RhlE在大肠杆菌内高效表达,动态光散射(DLS)分析显示其主要以单体的形态分布在溶液中.在ATP/Mg离子的辅助下可以高效解旋双链RNA,小角散射实验表明RhlE具有与同源家族蛋白类似的三维结构.更多还原

【Abstract】 Tuberculosis caused by Mycobacterium tuberculosis is one of the most serious and infectious chronic infectious diseases in the world.It has the characteristics of latent infection.RhlE,a RNA helicase derived from Mycobacterium tuberculosis,depends on the ATP hydrolysis activity,participates in RNA metabolism and regulates the assembly and degradation of ribosome.However,its catalytic mechanism and structural basis are still unclear.In this study,RhlE was expressed and purified in vitro,and its activity was determined by dynamic light scattering(DLS).The results showed that RhlE expressed well in E.coli.DLS analysis showed that RhlE was mainly distributed as monomer in the solution.Double-stranded RNA can be efficiently unscrewed with the assistance of ATP/Mg ions by RhlE.Small angle scattering experiments showed that RhlE had a threedimensional structure similar to that of homologous family proteins.更多还原