烷烃羟化酶基因alkB表达载体的构建及其在大肠杆菌中的诱导表达  

Construction and induced expression of alkB expression vector in Escherichia coli

作　　者：孙敏[1] 罗群[1] 何颖[1] 侯登勇[1] 王庆蓉[1] 沈先荣[1] 

机构地区：[1]海军医学研究所,上海200433

出　　处：《中华航海医学与高气压医学杂志》2013年第1期50-53,共4页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

摘　　要：目的构建烷烃羟化酶基因alkB表达载体并检测其在大肠杆菌的表达。方法体外合成alkB基因，选择烷烃表达载体pCom8，构建含alkB基因的重组质粒pCom8-alkB，并采用CaCl，法将其转入大肠杆菌DH5α中，通过十二烷基碘酸钠-聚丙烯酰胺凝胶电泳（SDS—PAGE）检测不同诱导条件下的alkB基因表达的变化。结果构建的含alkB基因的重组质粒在DH5α中得到有效表达，且随诱导时问的延长而增加，在柴油诱导浓度为2％～3％（Ⅴ/Ⅴ）时，目的蛋白表达量相对较大。结论成功构建了含alkB基因的表达载体，阐明r不同诱导条件对目的蛋白表达的影响规律。Objective To construct alkB-expressing vector and investigate alkB protein expression in Escherichia coll. Methods The alkane hydroxylase （alkB） gene was synthesized in vitro. An alkB-expressing vector, pCom8-alkB, was constructed and then transformed into E. coli, DH5α by using CaCI2. Changes in the expression of alkB were detected by SDS-PAGE in E. coli, DH5α by induction with diesel oil. Results SDS- PAGE analysis demonstrated that alkB was successfully expressed after induction. The expression levels of alkB were highest, when alkB expression was induced by adding diesel oil into the culture at the final concentration of 2% -3% （v/v） , and the expression levels increased with the increase of induction time. Conclusions The alkB-expressing vector was successfully constructed, and the regular pattern for the expression of target protein under different induction conditions was clearly explained.

关 键 词：烷烃羟化酶 表达载体 诱导表达 

分 类 号：Q[生物学]