携带多克隆位点的链霉菌-大肠杆菌穿梭载体pRES18和pRES19的构建。

Construction of pRES18 and pRES19, Streptomyces-Escherichia coli shuttle vectors carrying multiple cloning sites.

作者信息

Ishikawa J, Niino Y, Hotta K

机构信息

Department of Bioactive Molecules, National Institute of Health, Tokyo, Japan.

出版信息

FEMS Microbiol Lett. 1996 Nov 15;145(1):113-6. doi: 10.1016/0378-1097(96)00397-7.

DOI:10.1016/0378-1097(96)00397-7

PMID:8931335

Abstract

We developed two Streptomyces-Escherichia coli shuttle vectors. The plasmid pRES102, consisting of the essential region of pRES1 and the thiostrepton resistance gene (tsr) fragment of pIJ702, was combined with the E. coli plasmid vector pUC18 or pUC19. The resulting shuttle vectors, designated pRES18 and pRES19, respectively, have relatively compact size (6.25 kb), low copy number, multiple cloning sites reciprocally arranged in opposite directions, and selection markers for both Streptomyces (tsr) and E. coli (beta-lactamase (bla) and beta-galactosidase (lacZ). These shuttle vectors are capable of carrying DNA fragments as long as 10 kb, of being maintained in S. griseus, S. lavendulae and S. lividans, and are compatible with pIJ702.

摘要

我们构建了两种链霉菌-大肠杆菌穿梭载体。质粒pRES102由pRES1的必需区域和pIJ702的硫链丝菌素抗性基因（tsr）片段组成，它与大肠杆菌质粒载体pUC18或pUC19相结合。由此产生的穿梭载体分别命名为pRES18和pRES19，它们具有相对紧凑的大小（6.25 kb）、低拷贝数、相互反向排列的多克隆位点，以及针对链霉菌（tsr）和大肠杆菌（β-内酰胺酶（bla）和β-半乳糖苷酶（lacZ））的选择标记。这些穿梭载体能够携带长达10 kb的DNA片段，可在灰色链霉菌、淡紫色链霉菌和变铅青链霉菌中维持，并且与pIJ702兼容。