丙酮酸脱羧酶基因和乙醇脱氢酶基因表达载体的构建  

Construction of Expression Vector of Pyruvate Decarboxylase and Alcohol Dehydrogenase Ⅱ Genes

作　　者：辛亮[1] 秦锐[1] 宋刚[1] 由田[1] 平文祥[1] 葛菁萍[1] 

机构地区：[1]黑龙江大学生命科学学院微生物黑龙江省高校重点实验室,黑龙江哈尔滨150080

出　　处：《微生物学杂志》2012年第2期9-14,共6页Journal of Microbiology

基　　金：国家高技术研究发展计划(863计划;2007AA100702-6);国家自然科学基金(31070446);黑龙江省教育厅重点项目(11551z011);黑龙江省科技攻关重大项目(GA07B401-6);科技创新人才研究专项资金项目优秀学科带头人(RC2010XK002028);黑龙江大学高层次人才(创新团队)支持计划(Hdtd2010-17)

摘　　要：以运动发酵单胞菌(Zymomonas mobilis)的总DNA为模板,PCR扩增运动发酵单胞菌中的丙酮酸脱羧酶(Pyruvate decarboxylase,PDC)基因和乙醇脱氢酶Ⅱ(Alcohol dehydrogenaseⅡ,ADHⅡ)基因。将丙酮酸脱羧酶基因和含核糖体结合位点(RBS)的乙醇脱氢酶基因串联起来置于T7启动子控制下,构成多顺反子表达质粒pQR-PRA。经酶切和PCR验证,表达载体构建成功。将pQR-PRA转入大肠埃希菌(Escherichia coli)BL21中,转化子的定性检测表明有酶表达,并且初步测定了2种酶的表达量。The genes of the pyruvate decarboxylase（PDC） and alcohol dehydrogenase II（ADHII） of Zymomonas mobilis（Zm） were amplified with PCR using total DNA of the Zm as a template.Then contacted the PDC gene with ADHII gene that contained ribosome synaptic site together and put them under the control of promoter T7 to construct poly-cistron expression plasmid pQR-PRA and confirmed with test and verification of restriction analysis and PCR.The pQR-PRA was transformed to E.coli BL21.And the qualitative detect indicated that the transformant had the enzyme expression and had initially determined the expression volume of the two enzymes.

关 键 词：运动发酵单胞菌 大肠埃希菌 丙酮酸脱羧酶 乙醇脱氢酶 载体构建 

分 类 号：Q933[生物学—微生物学]