[大肠杆菌丝氨酸羟甲基转移酶基因（glyA）的克隆与表达]

[Cloning and expression of the E. coli serine hydroxymethyltransferase gene (glyA)].

作者信息

Shen T, Na S, Yu G, Xie J, Jia P

机构信息

Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080.

出版信息

Wei Sheng Wu Xue Bao. 1997 Dec;37(6):423-8.

PMID:11189372

Abstract

The E. coli K12 glyA gene(13 kb), 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 (13 kb) into plasmids pBR322, pBR329 and pSMY901 established that a 2.6 kb PstI-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.

摘要

编码丝氨酸羟甲基转移酶（SHMT）的大肠杆菌K12 glyA基因（13 kb）已通过插入失活和互补试验克隆到质粒载体pBR329中。将原始插入片段（13 kb）的片段亚克隆到质粒pBR322、pBR329和pSMY901中，确定一个2.6 kb的PstI-EcoR片段携带glyA基因。获得了12株含有重组质粒的转化体。携带glyA质粒的菌株中SHMT和glyA基因产物水平不同。在宿主菌株中未观察到SHMT活性。JM109(pSM13)、K12(pSM13)、K12(pSM14)和K12(pSM15)的glyA基因产物分别占总可溶性细胞蛋白的15.7%、15.4%、11.8%和9.48%。