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首页> 外文期刊>New biotechnology >Simultaneous expression of the bacterial Dictyoglomus thermophilum xynB gene under three different trichoderma reesei promoters
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Simultaneous expression of the bacterial Dictyoglomus thermophilum xynB gene under three different trichoderma reesei promoters

机译：在三种不同的里氏木霉启动子下同时表达嗜热嗜热乳球菌xynB基因

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摘要

Multiple copies of expression cassettes driven by the Trichoderma reesei xylanase 2 (xyn2) and cellobiohydrolase 2 (cbh2) promoters were introduced into the recombinant T. reesei EC-21 generated to express a thermostable Dictyoglomus thermophilum xylanase (XynB) under the egl2 promoter for further improvement of the enzyme yield. The transformants were screened based on increased XynB activity only. Multiple promoter transformant MPP-4 expressing the xynB gene under all the three promoters was found to be the highest producer of XynB, giving a 65% increase in yield compared to the parental single-promoter recombinant EC-21. The multiple-promoter transformant strains harboured six to nine copies of the xynB gene. Amongst the three promoters, egl2 seemed to have the strongest effect on XynB expression. The shotgun approach we used proved to be effective for rapid enhancement of protein expression using three promoters active at the near-neutral pH of the cultivation medium.
机译：由里氏木霉木聚糖酶2（xyn2）和纤维二糖水解酶2（cbh2）启动子驱动的表达盒的多个副本被引入到重组里氏木霉EC-21中，该重组产生的表达物在egl2启动子下表达了热稳定的嗜热双歧球菌木聚糖酶（XynB），以进一步表达酶产量的提高。仅基于增加的XynB活性筛选转化体。发现在所有三个启动子下均表达xynB基因的多个启动子转化体MPP-4是XynB的最高产量，与亲本单启动子重组EC-21相比，产率提高了65％。多启动子转化株具有xynB基因的六到九个拷贝。在这三个启动子中，egl2似乎对XynB表达具有最强的作用。我们使用的shot弹枪方法被证明可以有效地使用三种在培养基中性pH值附近具有活性的启动子快速增强蛋白质表达。

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