赤红球菌JDM312环己酮单加氧酶的原核表达及检测

journal6 ›› 2011, Vol. 32 ›› Issue (4): 88-91.

赤红球菌JDM312环己酮单加氧酶的原核表达及检测

（吉首大学生物资源与环境科学学院，湖南吉首 416000）

出版日期:2011-07-25 发布日期:2012-04-07
通讯作者: 吾鲁木汗·那孜尔别克(1961-)，女，新疆塔城托里人，吉首大学生物资源与环境科学学院教授，博士，主要从事微生物学研究.E-mail:ulum@jsu.edu.cn.
基金资助:
湖南省大学生研究性学习和创新性实验计划项目(CX2008146)

Prokaryotic Expression and Detection of Cyclohexanone Monooxygenase from Rhodococcus ruber JDM312

(College of Biology and Environmental Sciences,Jishou University,Jishou 41600,China)

Online:2011-07-25 Published:2012-04-07

摘要/Abstract

摘要：通过酶切法从重组质粒pUC18-chnB中得到编码环己酮单加氧酶的chnB基因序列，将其定向插入原核表达载体pQE30中，构建重组质粒pQE30-chnB，转化到大肠杆菌BL21中并诱导表达目的蛋白，用SDS-PAGE电泳检测表达产物.测序结果表明chnB基因大小为1 623 bp，编码由540个氨基酸残基构成的多肽.SDS-PAGE结果显示，表达分子量约为60 kDa的带有6 × His标签的环己酮单加氧酶，与预期分子量相符，表明成功构建出原核表达质粒并实现了目的蛋白表达，为进一步开展环己酮单加氧酶活性研究奠定了基础.

关键词： 赤红球菌, 环己酮单加氧酶, chnB基因, 原核表达

Abstract: The chnB gene encoding a cyclohexanone monooxygenase was isolated from the recombinant plasmid pMD18-chnB by digested with BamHI and HindⅢ,then cloned into the expression vector pQE30 and expressed in E. coli BL21 by IPTG induction.The expressed protein was identified by SDS-PAGE.The sequence analyses showed that the coding region of the chnB of Rhodococcus ruber JDM312 was 1 623 bp in length,and the predicted primary protein was composed of 540 amino acids.The SDS-PAGE analyses revealed a single protein band with a molecular weight of 60 kDa,suggesting that the recombinant plasmid of pQE30-chnB was successfully constructed and the target protein was expressed in E. coli.

Key words: rhodococcus ruber, cyclohexanone monooxygenase, chnB gene, prokaryotic expression

彭哲慧, 龚凤娟, 刘丹丹, 吾鲁木杆.那孜尔别克. 赤红球菌JDM312环己酮单加氧酶的原核表达及检测[J]. journal6, 2011, 32(4): 88-91.

PENG Zhe-Hui, GONG Feng-Juan, LIU Dan-Dan, WU Lu-Mu-Gan-.Na-Zi-尔Bie-Ke. Prokaryotic Expression and Detection of Cyclohexanone Monooxygenase from Rhodococcus ruber JDM312[J]. journal6, 2011, 32(4): 88-91.

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