金龟子绿僵菌(Metarhizium anisopliae HN1)几丁质酶与谷胱甘肽S-转移酶GST在大肠杆菌中的高效融合表达High-efficient Fusion Expression of Chitinase and GST from Metarhizium anisopliae HN1 in Escheichia coli
任文彬;张世清;黄俊生;
摘要(Abstract):
使用RT-PCR方法,从高毒力金龟子绿僵菌Metarhizium anisopliaeHN1中,克隆得到一个全长为1 275 bp的几丁质酶基因,经Blast分析此基因序列与M.anisopliaeE6的chi1基因(AF02749)同源率为96%。将此基因克隆到pGEX-6p-1载体上,使之与载体上一个约26kD大小的谷胱甘肽S-转移酶(GST)相连,构建pGEX-chi融合表达载体,转化到大肠杆菌(Escherichia coli)BL 21中,经SDS-PAGE结果分析显示:表达出的融合蛋白大小为68 kD,此目的蛋白占表达总量的64.5%。经破碎处理后可检测到几丁质酶活性。
关键词(KeyWords): 金龟子绿僵菌;几丁质酶基因;融合蛋白;大肠杆菌
基金项目(Foundation): 国家科技支撑计划(2007BAD48B00)
作者(Author): 任文彬;张世清;黄俊生;
Email:
DOI: 10.13989/j.cnki.0517-6611.2009.17.077
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