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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Construction of a plasmid for heterologous protein expression with a constitutive promoter in Trichoderma reesei
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Construction of a plasmid for heterologous protein expression with a constitutive promoter in Trichoderma reesei

机译:里氏木霉中具有组成型启动子的异源蛋白表达质粒的构建

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摘要

Trichoderma reesei is widely used as a host for homologus and heterologus protein expression because of its well-known capability of protein secretion, especially cellulases secretion. In this study, a binary vector which could be used for protein expression was constructed with a constitutive promoter of rp2 gene. This vector contained the expression cassette Prp2-target gene-Trp2 and hygromycin B selection marker based on pCAMBIA1300 for T-DNA random insertion. The feasibility of the promoter was determined by eGFP (enhanced green fluorescence protein) expression in T. reesei. For heterologus protein expression, the expression of bgla from Aspergillus niger in the transformant was 3 folds higher than that of the wild type strain. The results demonstrate that this constitutive promoter could be applied for protein expression and thus this protein express system may contribute to the industrial protein production.
机译:里氏木霉由于其众所周知的蛋白质分泌能力,尤其是纤维素酶的分泌能力,被广泛用作同源和异源蛋白质表达的宿主。在这项研究中,用rp2基因的组成型启动子构建了可用于蛋白质表达的二元载体。该载体包含表达盒Prp2-靶基因-Trp2和基于pCAMBIA1300的潮霉素B选择标记,用于T-DNA随机插入。启动子的可行性由里氏木霉中的eGFP(增强型绿色荧光蛋白)表达确定。对于异源蛋白表达,来自黑曲霉的bgla在转化体中的表达比野生型菌株高3倍。结果表明该组成型启动子可用于蛋白质表达,因此该蛋白质表达系统可能有助于工业化蛋白质的生产。

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