结核分枝杆菌基因敲除技术的研究进展

Reviews of gene knockout technology in Mycobacterium tuberculosis

结核分枝杆菌是十分重要的人兽共患病原菌,基因缺失株的构建在该菌致病与免疫机制、基因功能解析和防控技术中意义重大,而该菌特有的复杂结构和生理生化特性也给缺失株的构建带来难度,为此有关结核分枝杆菌基因敲除技术的研究一直是重要课题。本文重点介绍了结核分枝杆菌基于特异性转导噬菌体、线性DNA片段、反选择标记、CRISPR技术以及ORBIT系统的基因敲除方法,同时对分枝杆菌的分子遗传操作发展进行了展望。

Mycobacterium tuberculosisis is an extremely important pathogenic bacterium,and the construction of mycobacterial mutants is highly important for assessing its pathogenicity and immune mechanisms,gene function,and prevention and control technologies.Genetic methods including mutation technologies in mycobacteria are difficult to perform,owing to the bacterium’s complex physiological characteristics and mechanisms.Therefore,research on gene mutations in mycobacteria has long been an important issue.For better understanding of current methods,targeted gene deletion methods in mycobacteria based on a specialized transduction method,linear DNA fragment method,anti-selection marker method,CRISPR technology and the ORBIT system are summarized,and the development of molecular genetic manipulation of Mycobacterium is also described.