用穿梭质粒pHR106及其重组衍生物对拜氏梭菌NRRL B - 592进行电转化。

Electro-transformation of Clostridium beijerinckii NRRL B-592 with shuttle plasmid pHR106 and recombinant derivatives.

作者信息

Birrer G A, Chesbro W R, Zsigray R M

机构信息

Department of Microbiology, University of New Hampshire, Durham 03824.

出版信息

Appl Microbiol Biotechnol. 1994 Mar;41(1):32-8. doi: 10.1007/BF00166078.

DOI:10.1007/BF00166078

PMID:7764636

Abstract

Conditions for transformation of the solventogenic anaerobe Clostridium beijerinckii NRRL B-592 with plasmid DNA via electroporation are described. Shuttle plasmid pHR106 and two derivatives constructed in this study were transferred and were expressed in this organism. One recombinant derivative of pHR106 was constructed by separately subcloning the clostridial tetracycline (tetP) resistance genes into pHR106. The second vector conferring erythromycin resistance was obtained via in-vivo recombination. The new constructs, termed pRZL and pRZE respectively, were then transferred to C. beijerinckii in order to evaluate their potential as shuttle vectors. The recombinant plasmids were shown to transfer to C. beijerinckii and were expressed as autonomously replicating vectors. The use of these plasmids as cloning and shuttle vectors for C. beijerinckii is discussed.

摘要

描述了通过电穿孔法用质粒DNA转化产溶剂厌氧微生物拜氏梭菌（Clostridium beijerinckii）NRRL B - 592的条件。穿梭质粒pHR106以及本研究构建的两个衍生物被转移并在该生物体中表达。pHR106的一个重组衍生物是通过将梭菌四环素（tetP）抗性基因分别亚克隆到pHR106中构建而成。第二个赋予红霉素抗性的载体是通过体内重组获得的。然后将分别称为pRZL和pRZE的新构建体转移到拜氏梭菌中，以评估它们作为穿梭载体的潜力。重组质粒被证明可转移到拜氏梭菌中，并作为自主复制载体表达。讨论了将这些质粒用作拜氏梭菌的克隆和穿梭载体的用途。