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米黑根毛霉脂肪酶基因的克隆表达及活性分析
苗长林1; 罗文1; 刘姝娜1; 吕鹏梅1; 李惠文1; 杨玲梅1; 袁振宏1; 蒋剑春2
2013
Source Publication生物技术通报
ISSN1002-5464
Issue6Pages:122
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。
Language英语
Document Type期刊论文
Identifierhttp://ir.giec.ac.cn/handle/344007/20623
Collection中国科学院广州能源研究所
Affiliation1.中国科学院广州能源研究所
2.中国林业科学院林产化学工业研究所
First Author AffilicationGuangZhou Institute of Energy Conversion,Chinese Academy of Sciences
Recommended Citation
GB/T 7714
苗长林,罗文,刘姝娜,等. 米黑根毛霉脂肪酶基因的克隆表达及活性分析[J]. 生物技术通报,2013(6):122.
APA 苗长林.,罗文.,刘姝娜.,吕鹏梅.,李惠文.,...&蒋剑春.(2013).米黑根毛霉脂肪酶基因的克隆表达及活性分析.生物技术通报(6),122.
MLA 苗长林,et al."米黑根毛霉脂肪酶基因的克隆表达及活性分析".生物技术通报 .6(2013):122.
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