人参皂苷前体工程酵母的构建
Jilin Daxue Xuebao (Gongxueban)/Journal of Jilin University (Engineering and Technology Edition)(2011)
Abstract
从人参发根中通过RT-PCR扩增了人参达玛烯二醇合成酶基因。并利用这一基因和酵母表达载体pAUR123构建了产达玛烯二醇的转基因酿酒酵母。阳性克隆经短梗霉素A抗性粗筛,并经PCR、SDS-PAGE和TLC验证。研究结果表明,这一酵母能够表达人参达玛烯二醇合成酶,并产生达玛烯二醇。达玛烯二醇合成酶在整个酵母生长期内均有表达。达玛烯二醇合成酶活和达玛烯二醇水平在YDP液体培养基中培养36 h和42 h时达到最高,分别为132U/g干细胞和251μg/L。
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Key words
Bioengineering,Dammarenediol synthase,Panax ginseng hairy roots,Saccharomyces cerevisiae
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