黑曲霉电转化条件的研究

Transformation of Aspergillus niger by Electroporation-Mediated Transformation

研究避免了繁琐的原生体制备过程,直接使用萌发的黑曲霉孢子进行电转化,以潮霉素B作为筛选标记,从孢子萌发时间、电场强度及质粒浓度等方面考察了电转化效率的影响因素。研究表明,针对A.nigerMGG029-ΔaamA,其理想的电转化条件:孢子龄为4d,孢子萌发时间为2h,电场强度为5kV/cm。在上述条件下分别使用1μg环状或线状pBC-Hygro质粒DNA进行转化,平均可以得到34个和51个转化子,而在同样条件下使用质粒pRS303H平均可以获得163个和258个转化子。

This study avoided the complex technological process of protoplast preparation and directly transformed the germinated spores of Aspergillus niger by electroporation-mediated transformation （EMT） method with hygromycin B as the screening marker.The effects of germination time,electric field intensity （EFI） and plasmid concentration on the transformation efficiency were examined.The study indicated that the appropriate EMT conditions for A.niger MGG029-aamA were：spore stadium was 4 d,2 h after germination was preferred and EFI at 5 kV/cm was the best.Under these conditions,34 and 51 transformants in average were obtained when 1 μg circular or linearized plasmid DNA of pBC-Hygro was used,respectively,and 163 and 258 transformants in average were obtained when plasmid DNA of pRS303H was used in the same way.