霉酚酸产生菌原生质体转基因方法的建立

Establishment of a genetic transformation system for Penicillium brevicompactum to produce mycophenolic acid

【目的】建立霉酚酸产生菌短密青霉菌的遗传转化体系。【方法】以腐草霉素抗性为选择标记,利用聚乙二醇介导原生质体融合,进行外源基因转化。【结果】聚乙二醇介导的短密青霉菌原生质体转化效率为每微克DNA 2-3个转化子;转化子的PCR检测结果显示外源基因已经整合到短密青霉菌基因组中,转化子抗性稳定。【结论】霉酚酸产生菌短密青霉菌转基因体系的建立为该菌进行分子生物学研究以及基因工程育种奠定了基础。

[ Objective ] We established a genetic transformation system for Penicilliurn brevicornpactum to produce mycophenolic acid. [ Methods] We developed protoplast transformation methods mediated by Polyethylene glycol, using phleomycin resistance gene （Sh ble） as a dominant selection marker. [ Result] The frequency of transformation was up to 2 - 3 transformants per ~g DNA ; analysis of the transformants by PCR showed that the foreign DNA had been integrated into the host genome. The transformants retained stable after generation. [ Conclusion] The establishment of the genetic transformation system of PeniciUium brevicompacttum could serve as the basis for the research of molecular biology and the breeding of gene engineering of the fungus.