大肠杆菌yqhD基因的克隆与表达  

Cloning and expression of putative oxidoreductase(YqhD) gene of Escherichia coli

作　　者：韦旭钦[1] 韦宇拓[1] 黄日波[1] 

机构地区：[1]广西大学生命科学与技术学院,广西南宁530005

出　　处：《广西农业生物科学》2005年第4期299-303,共5页Journal of Guangxi Agricultural and Biological Science

基　　金：国家863计划项目(2003AA001039)

摘　　要：以大肠杆菌E scherich ia coli K-12基因组DNA为模板,利用PCR技术扩增得到假定的氧化还原酶(pu tative ox idoreductase)基因yqhD,将它连接到克隆质粒pGEM-3zf(+)上,得到重组质粒pGEM-yqhD,对此重组质粒进行序列测定,对其DNA序列分析表明,yqhD基因全长为1 164 bp。再将yqhD基因插入表达载体pSE 380,构建成重组子pSE 380-yqhD,并在E.coli BL 21中获得表达。研究表明,以1,3-丙二醇为底物时,基因工程菌在30°C下,以1.0 mm o l/L IPTG诱导12 h的酶活力达到3.13 U/mL,比对照菌株提高4.4倍。The yqhD gene encoding putative oxidoreductase （YqhD） was amplified from the genome of Escherichia coli strain K-12 by polymerase chain reaction（PCR） and subcloned into pGEM- 3zf（＋） to obtain recombinant plasmids pGEM-yqhD. It was shown from DNA sequence analysis that the open reading frame of yqhD was 1 164 bp. The gene of yqhD was inserted into expression vector pSE380 and then transformed into E. coli BL21. The results showed that the enzymatic activity of YqhD in crude extracts reached up to 3.13 U/mL when using 1, 3-propanediol as substrate under the induced condition of 1.0 mmol/L IPTG at 30℃ for 12 hours, and it was increased 4.4-fold compared with that of host strain.

关 键 词：大肠杆菌 假定的氧化还原酶 yqhD基因 克隆与表达 

分 类 号：Q78[生物学—分子生物学]