纤细裸藻△9-脂肪酸延长酶基因在解脂耶氏酵母中的表达

Expression of Euglena gracilis △9 fatty acid elongase gene in Yarrowa Lipolytica

摘要: 从纤细裸藻c DNA中克隆出△9-脂肪酸延长酶基因,与解脂耶氏酵母整合载体p INA1297连接构建重组质粒p INA1297-△9E,用电击转化法将重组质粒转化到解脂耶氏酵母中,筛选得到阳性表达菌株YL△9E。在酵母浸出粉胨葡萄糖培养（YPD）条件下,工程酵母YL△9E的菌体生长速率高于对照菌株polf。对工程酵母YL△9E进行脂肪酸成分分析,结果表明△9-脂肪酸延长酶基因获得表达,产生了二十碳二烯酸（EDA）,其含量可高达14.8%。

Abstract: The △9-fatty acid elongase was cloned from c DNA of the Euglena gracilis. The sequence from c DNA was ligated into p INA1297,yielding the recombinant p INA1297-△9E. Yarrowa Lipolytica Po1 f was transformed with p INA1297- △9E by electroporation method and the positive transformants YL△9E were obtained by YNBcasa plates. The growth rate of transgenic strains was much higher than that of wild strain in YPD medium.Analysis of gas chromatography/mass spectrometer(GC-MS) showed that eicosadienoic acid(EDA;C20∶2n-6)was detected and their percentage to total fatty acids in transgenic Yarrowa Lipolytica was 14.8%,which indicated that Euglena gracilis △9-fatty acid elongase gene was expressed in transgenic Yarrowa Lipolytica.