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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Heterologous protein expression in Trichoderma reesei using the cbhII promoter
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Heterologous protein expression in Trichoderma reesei using the cbhII promoter

机译：使用cbhII启动子在里氏木霉中异源蛋白表达

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摘要

To express homologous or heterologous proteins in fungi, a protein expression system using the promoter of cellobiohydrolase II gene (cbhII) was constructed by generating an expression vector called pWEIIF00. The obtained vector possesses the left and right borders, a hygromycin phosphotransferase B selective marker and a strong promoter and terminator of cbhII from Trichoderma reesei. It can easily undergo random recombination. The applicability of the vector was tested by red fluorescent protein gene (DsRed2) expression detection in T. reesei Rut C30. Using this system, a recombinant Cel5A variant, N342R (Qin et al., 2008), was then selected to express in Rut-C30. Compared to that of the parent strain, integration of the N342R gene resulted in 31.09% increased carboxymethyl-cellulose-degrading (CMCase) activity at pH 5.0 and 56.06% increased activity at pH 6.0. The increased CMCase activity of the recombinant strains would be beneficial for its application uses in multiple industries. The vector constructed in this study can used in fungi to produce industrial proteins.
机译：为了在真菌中表达同源或异源蛋白质，通过产生称为pWEIIF00的表达载体来构建使用纤维二糖水解酶II基因（cbhII）的启动子的蛋白质表达系统。所获得的载体具有左右边界，潮霉素磷酸转移酶B选择标记以及来自里氏木霉的cbhII的强启动子和终止子。它可以很容易地进行随机重组。通过在里氏木霉Rut C30中的红色荧光蛋白基因（DsRed2）表达检测来检验该载体的适用性。然后，使用该系统，选择重组Cel5A变体N342R（Qin等，2008）在Rut-C30中表达。与亲本菌株相比，N342R基因的整合导致pH 5.0时羧甲基纤维素降解（CMCase）活性提高31.09％，pH 6.0时活性提高56.06％。重组菌株CMCase活性的提高将有利于其在多个行业中的应用。本研究中构建的载体可用于真菌生产工业蛋白质。

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