大肠杆菌丝氨酸羟甲基转移酶基因(glyA)的克隆和表达  

CLONING AND EXPRESSION OF THE E. COLI SERINE HYDROXYMETHYLTRANSFERASE GENE (glyA)

作　　者：沈天翔[1] 那淑敏[1] 喻国策 谢家仪[1] 贾盘兴[1] 

机构地区：[1]中国科学院微生物研究所,北京100080

出　　处：《微生物学报》1997年第6期423-428,共6页Acta Microbiologica Sinica

基　　金：国家"八五"攻关项目

摘　　要：利用插入失活及营养缺陷型互补法将大肠杆菌K12 13kb的glyA基因克隆到质粒pBR329中。将重组质粒酶切,亚克隆,确定2.6kb PstI-EcoRI亚克隆片段带有完整的glyA基因。共获得12株glyA基因重组菌,对重组质粒进行了酶切鉴定。不同重组菌丝氨酸羟甲基转移酶(SHMT)活性及其酶表达量均不相同。受体菌未检测到丝氨酸的产生。重组菌株JM109(pSM13)、K12(pSM13)、K12(pSM14)和K12(pSM15)SHMT酶表达量分别占全菌可溶性蛋白的15.7％、15.4％、11.8％和9.48％。The E. coli K12 glyA gene(13kb), encoding serine hydroxymethyltransferase (SHMT), has been cloned in the plasmid vector pBR329 using insertion inactivation and complementation test. Subcloning of segments of the original insert (13kb) into plasmids pBR322, pBR329 and pSMY901 established that a 2.6kb Pstl-EcoR fragment carries the glyA gene. The 12 strains of transforments containing recombined plasmid. were obtained. SHMT and glyA gene product level in strains carrying glyA plasmids were different. No SHMT activity was observed in host strains. The glyA gene products for JM109(pSM13), K12(pSM13), K12(pSM14) and K12(pSM15) account for 15.7%, 15.4%, 11.8%, and 9.48% of the total dissoluble cell protein, respectively.

关 键 词：大肠杆菌 丝氨酸 羟甲基转移酶 glyA基因 克隆 

分 类 号：Q939.121[生物学—微生物学] Q78