TaqDNA聚合酶表达载体的构建及表达条件优化  

Construction of Taq DNA Polymerase Expression Vector and Optimization of Expression Conditions

作　　者：鞠守勇[1] JU Shou-yong(School of Bioengineering,Wuhan Polytechnic,Wuhan 430074,China)

机构地区：[1]武汉职业技术学院生物工程学院,湖北武汉430074

出　　处：《武汉职业技术学院学报》2022年第5期106-109,共4页Journal of Wuhan Polytechnic

基　　金：2021年湖北省教育厅科学技术研究指导性项目(B类项目)“核酸检测用快速Taq DNA聚合酶的研制”(项目编号:B2021467);2021年武汉职业技术学院校级重点项目“核酸检测用Taq DNA聚合酶的研制”(项目编号:2021YK010)。

摘　　要：Taq DNA聚合酶是基因工程中最重要的商业化工具酶之一,对于大量分子克隆的教学实验来说,存在一定的经济成本负担。一般基因工程实验室都有能力自主生产Taq酶,这样可以节省大量的科研经费,同时还可以培养学生综合实训能力。研究构建Taq DNA聚合酶表达载体,在大肠杆菌E.coli BL21(ED3)中表达,并成功提取Taq酶,检测其活性。PCR实验表明研究得到的Taq酶与商业化Taq酶的活力相当,为自主生产Taq DNA聚合酶教学实验室和研究实验室提供参考,极大降低实验室Taq酶的成本。Taq DNA polymerase is one of the most important commercial tool enzymes in genetic engineering.There is a certain economic cost burden for the teaching experiment of a large number of molecular cloning.General genetic engineering laboratories have the ability to independently produce Taq enzyme,which can save a lot of scientific research funds and cultivate students’comprehensive practical training ability.In this study,Taq DNA polymerase expression vector was constructed and expressed in E.coli BL21(ED3).Taq enzyme was successfully extracted and its activity was examined.The PCR experiments showed that the activity of Taq enzyme in this study is equivalent to that of commercial Taq enzyme,which provides an important reference for the teaching laboratory and research laboratory of independent production of Taq DNA polymerase,,thus greatly reducing the cost of laboratory Taq enzyme.

关 键 词：Taq DNA聚合酶 基因工程 Taq DNA聚合酶表达 

分 类 号：Q55[生物学—生物化学] TQ28[化学工程—有机化工]