高山被孢霉原生质体转化及延长酶基因同源表达的研究

Study on Protoplast Transformation of Mortierella alpina and Its Homologous Expression of Elongase Gene

为了有效提高高山被孢霉(Mortierella alpina)产花生四烯酸的能力,采用聚乙二醇介导的原生质体转化法将重组pD4质粒(含高山被孢霉合成花生四烯酸的延长酶基因GLELO)转入高山被孢霉中,对其转化条件和同源表达进行了研究.结果表明,原生质体最佳转化条件为:以山梨醇作为稳定剂,50mmol/L的Ca2+和聚乙二醇4000作为转化介质,于25℃转化15min后温育,原生质体转化率最高为1.8μg-1.该转化子的摇瓶实验结果显示重组菌中花生四烯酸的产量比原始菌株的产量最高提高了近30%.该研究结果为高山被孢霉的性状改进及其他真菌的的遗传转化提供参考.

In order to improve the production of Arachidonic acid （ARA）, the protoplasts from Mortierella alpina were transformed with recombinant pD4 plasmid carrying the GLELO gene （elongase gene） in PEG system to study its homologous expression. The results showed that the best transformation system for preparing protoplasts sorbitol as osmotic stabilizer with 50 mmol/L Ca2＋ and PEG4000 as the transformant medium,which was incubated at 25 ℃ for 15 min. After regenerating,the highest transformation ratio of 1.8 μg-1 was got. The results in shaking culture showed that ARA yield was almost improved by 30% in the recombinant Mortierella alpina than the original strain. It＇s suggested that it will be useful for studying on the improvements of traits in Mortierella alpina and the genetic transformation in other fungi.