一系列用于枯草芽孢杆菌的整合质粒，在所有三个开放阅读框中都含有用于翻译 lacZ 融合的独特克隆位点。

A series of integrative plasmids for Bacillus subtilis containing unique cloning sites in all three open reading frames for translational lacZ fusions.

作者信息

Dahl M K, Meinhof C G

机构信息

Institut für Mikrobiologie und Biochemie, Universität Erlangen, Nürnberg, Germany.

出版信息

Gene. 1994 Jul 22;145(1):151-2. doi: 10.1016/0378-1119(94)90341-7.

DOI:10.1016/0378-1119(94)90341-7

Abstract

Two sets of three plasmids each were constructed based on integrative plasmids for Bacillus subtilis. Each set encodes resistance to either chloramphenicol or kanamycin. The plasmids contain a cassette consisting of the resistance gene, BamHI and SmaI cloning sites, and the promoterless lacZ gene. The cassette is flanked by the 3' and 5' ends of the amyE gene (encoding amylase) allowing integration of the cassette into that locus in the B. subtilis chromosome. For propagation and selection in Escherichia coli, the plasmids contain the pBR322 origin of replication and the beta-lactamase-encoding gene. Within a set, each of the three plasmids carries the unique SmaI and BamHI restriction sites in a different reading frame relative to the 'lacZ gene. These sets of plasmids allow the easy construction of translational fusions with lacZ and single-copy integration at the amyE locus of B. subtilis.

摘要

基于枯草芽孢杆菌整合质粒构建了两组质粒，每组三个。每组分别编码对氯霉素或卡那霉素的抗性。这些质粒包含一个盒式结构，由抗性基因、BamHI和SmaI克隆位点以及无启动子的lacZ基因组成。该盒式结构两侧是amyE基因（编码淀粉酶）的3'和5'末端，可使盒式结构整合到枯草芽孢杆菌染色体的该位点。为了在大肠杆菌中进行增殖和筛选，质粒含有pBR322复制起点和β-内酰胺酶编码基因。在一组质粒中，三个质粒中的每一个相对于“lacZ基因都以不同的阅读框携带独特的SmaI和BamHI限制性位点。这些质粒组便于构建与lacZ的翻译融合体，并在枯草芽孢杆菌的amyE位点进行单拷贝整合。