解纤维梭菌两个纤维素酶基因在大肠杆菌中的克隆与表达

Cloning and expression of two cellulase genes of Clostridium cellulolyticum in Escherichia coli.

作者信息

Faure E, Bagnara C, Belaich A, Belaich J P

机构信息

Laboratoire de Chimie Bactérienne, Centre National de la Recherche Scientifique, Marseille, France.

出版信息

Gene. 1988 May 15;65(1):51-8. doi: 10.1016/0378-1119(88)90416-7.

DOI:10.1016/0378-1119(88)90416-7

PMID:2840356

Abstract

Two cellulase genes isolated from Clostridium cellulolyticum strain ATCC3519 were cloned in Escherichia coli using plasmid pACYC184. Plasmids pB52 and pB43 were isolated from the transformants producing carboxymethylcellulase (CMCase) and the two cloned CMCase-coding genes were found to be included in two EcoRI fragments of 5.7 kb and 2.6 kb, respectively. These two genes showed no homology. The CMCase-coding genes were found to be contained in a 1.8-kb KpnI-HindIII fragment and a 2.05-kb HindIII-PvuII fragment of the DNA donor strain. Expression of these genes in E. coli was found not to depend on their orientation in the cloning vector. Hybridization experiments between these two fragments and Clostridium thermocellum NCIB10682 DNA fragments carrying genes celA, celB, celC and celD were carried out and some homologies were detected.

摘要

从解纤维素梭菌菌株ATCC3519中分离出的两个纤维素酶基因，利用质粒pACYC184克隆到大肠杆菌中。从产生羧甲基纤维素酶（CMCase）的转化子中分离出质粒pB52和pB43，发现两个克隆的CMCase编码基因分别包含在5.7 kb和2.6 kb的两个EcoRI片段中。这两个基因没有同源性。发现CMCase编码基因包含在DNA供体菌株的一个1.8 kb KpnI-HindIII片段和一个2.05 kb HindIII-PvuII片段中。发现这些基因在大肠杆菌中的表达不依赖于它们在克隆载体中的方向。对这两个片段与携带celA、celB、celC和celD基因的嗜热栖热菌NCIB10682 DNA片段进行了杂交实验，检测到了一些同源性。