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

作　　者：王海宁 金珊珊 徐正中 王蕾 焦新安 陈祥 WANG Hai-ning;JIN Shan-shan;XU Zheng-zhong;WANG Lei;JIAO Xin-an;CHEN Xiang(Jiangsu Key Laboratory of Zoonosis,Jiangsu Co-Innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses,Yangzhou University,Yangzhou 225009,China;Key Laboratory of Prevention and Control of Biological Hazard Factors(Animal Origin)for Agrifood Safety and Quality,Ministry of Agriculture and Rural Affairs,Yangzhou University,Yangzhou 225009,China)

机构地区：[1]扬州大学,江苏省人兽共患病学重点实验室/江苏省动物重要疫病与人兽共患病防控协同创新中心,扬州225009 [2]扬州大学,农业农村部农产品质量安全生物性危害因子(动物源)控制重点实验室,扬州225009

出　　处：《中国人兽共患病学报》2020年第11期928-933,共6页Chinese Journal of Zoonoses

基　　金：国家重点研发计划(2017YFD0500300);江苏省自然科学基金(BK20171285);江苏‘六大人才高峰’(SWYY-083);优势学科建设工程项目(PAPD)联合资助。

摘　　要：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.

关 键 词：结核分枝杆菌 基因敲除 特异性转导 线性DNA片段 反选择标记 CRISPR ORBIT 

分 类 号：R378.91[医药卫生—病原生物学]