Development of a versatile shuttle vector for gene expression in Geobacillus spp.
Development of a versatile shuttle vector for gene expression in Geobacillus spp.
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DOI：
10.1016/j.plasmid.2008.04.001
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发表时间：
2008-07-01
期刊：
影响因子：
2.6
通讯作者：
Leak, David J.
中科院分区：
文献类型：
--
作者：
Taylor, Mark P.;Esteban, Carlos D.;Leak, David J.
一个改进的，多功能的穿梭载体已被创建为土芽孢杆菌属的代谢工程。由于卡那霉素是最耐热的常用抗生素，编码耐热卡那霉素核苷酸转移酶的基因，连同来自G。将嗜热脂肪菌质粒pBST 1克隆到大肠杆菌克隆载体pUC 18中。所得到的载体命名为pUCG 18，在两种生物体中复制，并且可以以1 × 10（4）转化体/g G.热葡萄糖苷酶，并在抗生素选择下稳定至68 ° C。利用该质粒构建了棕榈发酵单胞菌丙酮酸脱羧酶（pdc）基因在G. pBST 1衍生的复制起点的序列分析显示与以前仅在巨大芽孢杆菌菌株中发现的θ复制子家族同源。(C)2008年爱思唯尔公司All rights reserved.
An improved, versatile shuttle vector has been created for the metabolic engineering of Geobacillus spp. As kanamycin is the most thermo-tolerant of commonly used antibiotics, the gene encoding a thermostable kanamycin nucleotidyltransferase, together with the origin of replication from the G. stearothermophilus plasmid pBST1 were cloned into the Escherichia coli cloning vector pUC18. The resulting vector, named pUCG18, replicated in both organisms and could be transformed with an efficiency of 1 x 10(4) transformants per mu g of DNA in G. thermoglucosidasius and was stable up to 68 degrees C with antibiotic selection. It was used to demonstrate expression of the pyruvate decarboxylase (pdc) gene from Zymomonas palmae in G. thermoglucosidasius at 45 degrees C.Sequence analysis of the pBST1 derived origin of replication revealed homology with a family of theta replicons that have previously only been found in strains of Bacillus megaterium. (C) 2008 Elsevier Inc. All rights reserved.