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

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%。

The △9-fatty acid elongase was cloned from cDNA of the Euglena gracilis. The sequence from cDNA was ligated into plNA1297,yielding the recombinant plNA1297-A9E. Yarrowa Lipolytica Polf was transformed with pINA1297-A9E by electroporation method and the poSitive transformants YLA9E 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 transqenic Yarrowa Lipolytica.