Establishment of a genetic transformation system in non-model fungus Penicillium raistrickii
Penicillium raistrickii can yield plenty of secondary metabolites.However,there are few reports about its synthetic pathways and unexpressed biosynthetic gene clusters because of the lack of a genetic transformation system,as a result,the study of its biosynthesis mechanism and natural products is seriously hampered.An efficient genetic transformation system for P.raistrickii CGMCC 3.1066 was built in this study.The conditions for spore germination,the antibiotic concentration,the optimum condition for preparing protoplasts,and the concentration of DNA fragments have been determined.Five genes ranging in length from 1-10 kb were knocked out by polyethylene glycol-mediated protoplast transformation,and the average positive rate of gene knockout was 17.5%.In addition,the gene for synthesizing 1,8-dihydroxy naphthalene(DHN)-melanin has been knocked out.The results show that the lack of this gene can cause spore discoloration from green to white,but the growth of the strain is not affected,making this site to act as a fixed insertion site because the transformants can be identified quickly through observation.The genetic transformation system successfully established in P.raistrickii CGMCC 3.1066 will facilitate the identification of gene function,analysis of biosynthetic pathways of compounds,and other studies,and this study can provide a reference for building the transformation system of other non-model fungi.
万方数据
维普
