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[嗜热栖热放线菌F7纤维素酶基因在大肠杆菌和枯草芽孢杆菌中的克隆与表达]

[Cloning and expression of Clostridium thermocellum F7 cellulase genes in Escherichia coli and Bacillus subtilis cells].

作者信息

Tsoĭ T V, Bukhtiiarova M G, Aminov R I, Golovchenko N P, Kataeva I A, Kosheleva I A, Akimenko V K, Boronin A M

出版信息

Genetika. 1990 Aug;26(8):1349-60.

PMID:2124195
Abstract

The Clostridium thermocellum total DNA Sau 3A fragments' library was constructed on the basis of shuttle vector pMK4 for the Escherichia coli - Bacillus subtilis. 14 clones with endoglucanase activity and one with beta-glucosidase activity were selected in E. coli cells. Recombinant plasmids pCE were characterized by structural instability of various degree in B. subtilis cells. The results of the physical mapping, analysis of gene products in E. coli mini-cells as well as the DNA-DNA blot hybridization have led to conclusion on cloning of 7 individual genes for endoglucanases. Up to 3 polypeptides of various molecular weight corresponding to the products of cel gene were revealed in E. coli mini-cells containing the recombinant plasmids. The hybridization analysis demonstrated considerable homology of the majority of cel genes.

摘要

基于大肠杆菌-枯草芽孢杆菌穿梭载体pMK4构建了嗜热栖热梭菌总DNA Sau 3A片段文库。在大肠杆菌细胞中筛选出14个具有内切葡聚糖酶活性的克隆和1个具有β-葡萄糖苷酶活性的克隆。重组质粒pCE在枯草芽孢杆菌细胞中表现出不同程度的结构不稳定性。物理图谱分析、大肠杆菌小细胞中基因产物分析以及DNA-DNA印迹杂交结果得出了7个内切葡聚糖酶个体基因被克隆的结论。在含有重组质粒的大肠杆菌小细胞中发现了多达3种对应于cel基因产物的不同分子量的多肽。杂交分析表明大多数cel基因具有相当高的同源性。

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