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首页> 外文期刊>Applied Microbiology and Biotechnology >Construction of protein overproducer strains in Bacillus subtilis by an integrative approach
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Construction of protein overproducer strains in Bacillus subtilis by an integrative approach

机译：整合法构建枯草芽孢杆菌蛋白质超生产菌株

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摘要

We evaluated the effect of several genetic factors reported as having a role in the induction of the expression of significant levels of recombinant protein in Bacillus subtilis. We utilized the beta -galactosidase reporter protein from Escherichia coli as our model for measuring the overproduction of heterologous proteins in B. subtilis. The lacZ gene was expressed in B. subtilis using the regulatory region of the subtilisin gene aprE. In this study, we considered factors known to modulate the transcription and translation initiation rates and genetic and mRNA stability. We also consider the effects of different genetic backgrounds, such as degU32 and hpr2, that until now have been studied independently. By changing the native -35 promoter box to the consensus TTGACA sequence of the aprE promoter, a significant 100-fold increase in the beta -galactosidase activity was obtained. On the other hand, changes such as the GTG to ATG start codon, the construction of a consensus AAGGAGG ribosome binding site, and the addition of the cryIIIA transcription terminator at the 3' end of the lacZ gene, produced only marginal effects on the final beta -galactosidase activity. [References: 21]
机译：我们评估了据报道在诱导枯草芽孢杆菌中显着水平的重组蛋白表达中起作用的几种遗传因素的作用。我们利用大肠杆菌的β-半乳糖苷酶报道蛋白作为我们的模型，以测量枯草芽孢杆菌中异源蛋白的过量生产。 lacZ基因在枯草芽孢杆菌中使用枯草杆菌蛋白酶基因aprE的调控区表达。在这项研究中，我们考虑了已知可调节转录和翻译起始速率以及遗传和mRNA稳定性的因素。我们还考虑了迄今为止尚未独立研究的不同遗传背景（例如degU32和hpr2）的影响。通过将天然的-35启动子框更改为aprE启动子的共有TTGACA序列，可以使β-半乳糖苷酶活性显着提高100倍。另一方面，从GTG到ATG的起始密码子的变化，共有的AAGGAGG核糖体结合位点的构建以及在lacZ基因3'末端添加cryIIIA转录终止子的变化，仅对最终序列产生边际效应β-半乳糖苷酶活性。 [参考：21]

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