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On-line Access: 2021-03-12
Received: 2020-09-04
Revision Accepted: 2020-11-16
Crosschecked: 2021-01-29
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Citations: Bibtex RefMan EndNote GB/T7714
Liujia SHI, Fangfang YANG, Yanyan XU, Shoufeng WANG. Expression of Drosophila melanogaster acetylcholinesterase (DmAChE) gene splice variants in Pichia pastoris and evaluation of its sensitivity to organophosphorus pesticides[J]. Journal of Zhejiang University Science B,in press.Frontiers of Information Technology & Electronic Engineering,in press.https://doi.org/10.1631/jzus.B2000525 @article{title="Expression of Drosophila melanogaster acetylcholinesterase (DmAChE) gene splice variants in Pichia pastoris and evaluation of its sensitivity to organophosphorus pesticides", %0 Journal Article TY - JOUR
果蝇乙酰胆碱酯酶基因剪接变种在毕赤酵母中的表达及其对有机磷农药敏感性的评价创新点:DmAChE-like是果蝇乙酰胆碱酯酶基因发生剪接突变产生的,在此之前并未被报道过。本文通过检测证明了DmAChE-like具有乙酰胆碱酯酶活性,并且对大多数有机磷农药敏感。这个发现为酶抑制法检测农药残留提供了新的酶源。 方法:在本研究中,我们通过构建毕赤酵母重组表达菌株GS115-pPIC9K-DmAChE-like并进行诱导表达,利用Ellman法检测其酶活,并进行酶学性质研究。我们选取了5种常见农药进行农药敏感性试验,比较其与DmAChE之间的差异性。 结论:在本研究中,我们构建了基因工程的GS115-pPIC9K-DmAChE-like菌株,并在实验室水平诱导了基因表达。我们测试了各个发酵上清液的酶活性,证明了DmAChE-like具有乙酰胆碱酯酶活性。通过Tricine-SDS-PAGE,我们发现特异的蛋白条带为4.3 kDa,与预期结果一致。农药敏感性试验发现DmAChE-like对大多数选择的有机磷农药敏感,表明它可以用作有机磷农药检测酶抑制方法的新酶源。 关键词组: Darkslateblue:Affiliate; Royal Blue:Author; Turquoise:Article
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