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通过异源质粒整合产生的罗克福特青霉蛋白酶缺陷菌株的特性:在蛋白质生产中的潜在用途。

Characterization of a protease deficient strain of Penicillium roqueforti generated by heterologous plasmid integration: potential use for protein production.

作者信息

Durand-Poussereau N, Fevre M

机构信息

LAboratoire de Biologie Cellulaire Fongique, Université Claude Bernard Bat 405, Villeurbanne, France.

出版信息

J Biotechnol. 1996 Oct 18;51(1):97-105. doi: 10.1016/0168-1656(96)01574-x.

Abstract

A strain of Penicillium roqueforti was transformed with a plasmid which confers resistance to phleomycin. Stable transformants were selected. They all present a high resistance to the antibiotic. Their proteolytic activity was tested. One transformant is a proteolytic deficient strain unable to degrade casein. It is characterized by a tandem integration of the transformant vector in one site of the genome. The extracellular proteins profile of the strain reveals the absence of a 43 kDa polypeptide which probably corresponds to the aspartyl protease of Penicillium roqueforti. The aspA gene which encodes aspartyl protease is not expressed in the transformant and Southern analyses show that the aspA gene is not disrupted by the transformation vector.

摘要

用赋予博来霉素抗性的质粒转化一株罗克福特青霉。选择出稳定的转化体。它们都对该抗生素表现出高抗性。测试了它们的蛋白水解活性。其中一个转化体是蛋白水解缺陷型菌株,无法降解酪蛋白。其特征是转化载体在基因组的一个位点串联整合。该菌株的细胞外蛋白质谱显示不存在一条43 kDa的多肽,该多肽可能对应于罗克福特青霉的天冬氨酰蛋白酶。编码天冬氨酰蛋白酶的aspA基因在该转化体中不表达,Southern分析表明aspA基因未被转化载体破坏。

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