荚膜红细菌转酮醇酶的分离：其结构基因tktA的克隆与测序

Isolation of Rhodobacter capsulatus transketolase: cloning and sequencing of its structural tktA gene.

作者信息

de Sury d'Aspremont R, Toussaint B, Vignais P M

机构信息

CEA Grenoble, Biochimie Microbienne, Département de Biologie Moléculaire et Structurale, France.

出版信息

Gene. 1996 Feb 22;169(1):81-4. doi: 10.1016/0378-1119(95)00796-2.

DOI:10.1016/0378-1119(95)00796-2

PMID:8635754

Abstract

Rhodobacter capsulatus transketolase (Tkt) protein has been isolated from strain B10 by heparin affinity chromatography. Oligodeoxyribonucleotides (oligo) constructed as based on the amino-acid sequences were used for polymerase chain reaction (PCR) amplification on total genomic DNA. Southern hybridization with the PCR product as a probe allowed the isolation of a 5-kb PstI DNA fragment containing the structural Tkt-encoding gene (tktA) which was cloned and sequenced. The deduced tktA product of 671 aa (72815 Da) shares 59% identity with Rhodobacter sphaeroides Tkt.

摘要

通过肝素亲和层析从荚膜红细菌B10菌株中分离出转酮醇酶（Tkt）蛋白。根据氨基酸序列构建的寡脱氧核糖核苷酸（oligo）用于对总基因组DNA进行聚合酶链反应（PCR）扩增。以PCR产物为探针进行Southern杂交，从而分离出一个包含编码Tkt的结构基因（tktA）的5 kb PstI DNA片段，并对其进行克隆和测序。推导的由671个氨基酸（72815 Da）组成的tktA产物与球形红细菌Tkt具有59%的同一性。