摘要
以运动发酵单胞菌的总DNA为模板,PCR扩增Zym om onas m obilis中的乙醇脱氢酶基因(adhB)和丙酮酸脱羧酶基因(p d c)。首先进行单个基因表达,基因表达产物经SDS-PAGE电泳分析和乙醛指示平板检测,确认有酶活性后,将这2个基因串联构建多顺反子表达质粒pSE-p d c-adhB,转入大肠杆菌DH 5α内,得重组工程菌株。工程菌株经IPTG诱导,分别在含10%葡萄糖和10%木糖培养基中于37℃培养72 h,有乙醇产生,酒精产率分别为对照菌株21倍和5倍。
In this study pdc and adhB genes were amplified by PCR from total DNA of Zymomonas rnobilis. Two genes were expressed separately in E. coli DH5α. Distinct bands were detected as the products of adhB and pdc on SDSPAGE. The specific activities were detected by aldehyde indicator plates, and then two genes were linked as an operon. The recombinant plasmid pSE-pdc-adhB was transformed into E. coli DH5α and induced to express ADH I and PDC with IPTG. Ethanol which recombinant strains produced from 10% glucose and 10% xylose was separately 21 times and 5 times than that of the the wild-type E. coli DH5α under the condition of 37℃ and 72 h.
出处
《广西农业生物科学》
CAS
CSCD
2005年第3期187-190,共4页
Journal of Guangxi Agricultural and Biological Science
基金
国家"863"资助项目(2001AA214171)
广西大学科研资金资助项目(X032013)
关键词
运动发酵单胞菌
丙酮酸脱羧酶
乙醇脱氢酶
克隆
诱导表达
Zymomonas mobilis
pyruvate decarboxylase
alcohol dehydrogenase
cloning
induce expression
