以CAT为报告基因的蜡样芽孢杆菌强组成型启动子的筛选及初步鉴定  

Screening and Identification of Constructive Promoters from the Ge- nome of Bacillus Cereus Using CAT as the Reporter Gene

作　　者：汪玉婷[1] 井申荣[1] 

机构地区：[1]昆明理工大学医学院,昆明市650500

出　　处：《医学分子生物学杂志》2017年第3期157-161,共5页Journal of Medical Molecular Biology

摘　　要：目的从蜡样芽孢杆菌基因组中筛选组成型启动子，并对其活性进行鉴定。方法用Sau3AI酶切蜡样芽孢杆菌的基因组，回收片段，连接载体pCMR8a，转化DH5ct感受态后涂布在氯霉素和卡那霉素的培养基上得到强启动子，并对其进行截短和顺反性实验，并检测其启动蛋白表达情况。结果成功筛选到一个具有反向启动外源蛋白表达能力的启动子序列，并能够有效表达多种外源蛋白。结论本实验成功筛选并验证了蜡样芽孢杆菌的一种组成型启动子，能够有效地启动外源蛋白的表达．对实验室中表达某种或某些蛋白和工业生产提供了更多的选择。Objective We screening the constitutive promoter from genome of the Bacillus cere- us, and detect their activity. Methods The genome DNA of Bacillus cereus was digested with re- striction endonuclease Sau3A I , recycling fragment, cloned into pCMR8a, the constructs were transformed into competent E. coli DH5α. the positive clones were screened by growing on the LB agar plate with both chloramphenicol and Kanamycin, to proceeding truncation and Cis-trans test, testing the ability of promoter above protein expression. Results successful screening to a se- quence, which the sequence have an ability that reverse activated exogenous protein expres- sion. Conclusion This experiment successfully screened and verified a Bacillus cereus constitutive promoter, expression of exogenous protein can effectively, provide more choices for the expression of some oroteins in the laboratorv and industrial oroduction.

关 键 词：蜡样芽孢杆菌 启动子 氯霉素乙酰转移酶 

分 类 号：R378.7[医药卫生—病原生物学]