中文摘要：

深黄被孢霉是国内研究生产γ-亚麻酸（γ-linolenic acid,GLA）和花生四烯酸（Arachidonic acid,AA）等多不饱和脂肪酸（Polyunsaturated fatty acid,PUFA）的主要产油丝状真菌.前期的实验结果表明,深黄被孢霉M6-22具有潮霉素抗性,而且目前也没有关于深黄被孢霉营养缺陷型菌株的报道,限制了一些基于深黄被孢霉菌株进行遗传操作的研究.研究以红色荧光蛋白DsRED基因作为报告基因,构建能同时用于丝状真菌外源基因和RNAi表达载体pS-DsRED.通过PEG/CaCl2原生质体转化法将pS-DsRED导入深黄被孢霉M6-22中进行表达,成功获得产粉红色的阳性菌落,并在此基础上构建了深黄被孢霉Δ^12脂肪酸脱氢酶基因RNAi表达质粒pSREDMID12RNAi,为下一步目的基因的敲除和基因功能分析奠定了基础.

英文摘要：

Mortierella isabellina is an oil-producing fungal species which has been widely studied for producing polyunsaturated fatty acids including γ-Linolenic acid and arachidonic acid in China.Our previous research showed that Mortierella isabellina M6-22 is a hygromycin resistant fungal strain,and no auxotrophic mutant strains from M.isabellina were constructed so far,which limits its applications of genetic manipulation.In this study,DsRed gene which encodes a red-fluorescent protein was used as a report gene to construct a vector for both heterologous filamentous fungal gene and RNAi expression.The resultant vector pS-DsRED was further delivered into M.isabellina M6-22 by PEG/CaCl2-mediated protoplast transformation for expression,and acquired pink positive colonies.Based on the pS-DsRED,an RNAi expression plasmid pSREDMID12RNAi targeting Monierella isabellina delta-12-fatty acid desaturase gene was constructed,which provided a basis for the subsequent target gene knockdown and gene functional analysis.