杜氏盐藻荧光素酶基因表达载体的构建  

Construction of expression plasmid vectors containing the luciferase gene of an algae Dunaliella salina

作　　者：柴玉荣[1] 王天云[1] 侯桂琴[1] 侯卫红[1] 谢华[1] 袁保梅[1] 王建民[1] 薛乐勋[1] 

机构地区：[1]郑州大学细胞生物学研究室,郑州450052

出　　处：《郑州大学学报（医学版）》2004年第1期29-31,共3页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省重大科技攻关基金资助项目 0 12 2 0 3 2 5 0 0 ;河南省杰出人才创新基金资助项目 0 2 2 10 0 190 0

摘　　要：目的 :构建盐藻荧光素酶基因表达载体。方法 :分别用合适的限制性内切酶消化载体pGL3-Enhancervector、pD -B、pMD -CA和pMD -rbcS ,胶回收适当的片段 ,T4DNA连接酶过夜连接 ,转化宿主菌大肠杆菌JM1 0 9。挑取阳性克隆 ,提取质粒DNA ,酶切鉴定。结果 :得到含盐藻自身启动子碳酸酐酶 (CA)基因启动子、双倍重复碳酸酐酶 (DCA)基因启动子和来自衣藻的 1 ,5二磷酸核酮糖羧化酶 /加氧酶小亚基 (rbcS)基因启动子 ,以及荧光素酶(Luc)基因为外源基因的 3个盐藻表达载体。结论 :构建了 3个杜氏盐藻荧光素酶表达载体。Aim: The purpose of this study was to construct expression vectors containing the heterogeneous gene of luciferase for Dunaliella salina. Methods: The plasmid vectors: pGL3-Enhancer vector, pD-B, pMD-CA and pMD-rbcS were digested, respectively, with suitable restriction endonucleases and the resulting fragments were extracted from agarose gel, and ligated by T4 DNA ligase. The recombination plasmids were transformed into E.coli JM109 and the positive clones were screened. Plasm DNA was extracted and identified. Results: Three expression vectors were constructed which contain promoter fragments of duplicated carbonic anhydrase (DCA), carbonic anhydrase (CA) genes from Dunaliella salina and ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) small subunit (rbcS) promoter from Chlamydomanos reinhardtii, as well as NospolyA.Conclusion:Three expression vectors containing the heterogeneous gene of luciferase for Dunaliella salina have been construced.

关 键 词：杜氏盐藻 荧光素酶基因 质粒表达载体 

分 类 号：Q782[生物学—分子生物学]