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米黑根毛霉脂肪酶基因的克隆、表达及活性分析
Authors
刘姝娜
吕鹏梅
+6 more
李惠文
杨玲梅
罗文
苗长林
蒋剑春
袁振宏
Publication date
1 January 2013
Publisher
Abstract
以米黑根毛霉(Rhizomucor miehei)为出发菌株,采用UNIQ-10柱式Trizol总RNA抽提试剂盒提取其总RNA,反转录-聚合酶链式反应(RT-PCR)扩增出米黑根毛霉脂肪酶(RML)结构基因。构建真核重组表达质粒RML-pPIC9K,电转化His+缺陷型巴斯德毕赤酵母(Pichia pastoris)GS115,利用MD-MM平板及PCR方法筛选和鉴定出重组子,进行甲醇诱导表达。结果表明,RML基因能够在巴斯德毕赤酵母中实现高效表达。重组子发酵液经SDS-PAGE分析显示获得了与预期大小(约39 kD)一致的蛋白表达特异条带,用NaOH滴定法测其活性,酶活可达84 U/mL
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Institutional Repository of GuangZhou Institute of Energy Conversion, CAS
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oai:ir.giec.ac.cn:344007/34089
Last time updated on 04/12/2021
Institutional Repository of GuangZhou Institute of Energy Conversion, CAS
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:ir.giec.ac.cn:344007/20623
Last time updated on 05/12/2019
Institutional Repository of GuangZhou Institute of Energy Conversion, CAS
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:ir.giec.ac.cn:344007/20621
Last time updated on 05/12/2019
Institutional Repository of GuangZhou Institute of Energy Conversion, CAS
See this paper in CORE
Go to the repository landing page
Download from data provider
oai:ir.giec.ac.cn:344007/34087
Last time updated on 04/12/2021