白藜芦醇合酶基因遗传转化杏鲍菇的研究  

Genetic Transformation of Pleurotus eryngii with Resveratrol Synthase Gene

作　　者：廖静文[1] 郭丽琼[1] 林俊芳[1] 游楚镇[1] 

机构地区：[1]华南农业大学食品学院,广东广州510642

出　　处：《生物技术》2013年第5期31-36,共6页Biotechnology

基　　金：国家自然科学基金项目("食用真菌组合表达白藜芦醇研究";31272217;"食用真菌组合表达紫杉醇及其中间产物的基础研究";31071837)资助~~

摘　　要：目的:研究利用杏鲍菇作为生物反应器表达白藜芦醇。方法:以杏鲍菇菌株Pe18和P811为材料,以潮霉素浓度梯度对杏鲍菇原生质体与菌丝体对潮霉素的敏感浓度进行测定。采用PEG介导法将白藜芦醇合酶基因(rs)和潮霉素抗性基因(hph)共转化进杏鲍菇的原生质体中,采用潮霉素筛选与PCR扩增对拟转化子进行筛选鉴定。结果:Pe18和P811菌株的原生质体对潮霉素的最低敏感浓度分别为20μg/mL和10μg/mL,两菌株的菌丝体对潮霉素的敏感性均为160μg/mL;90个拟转化子中有30个同时整合了两个基因。结论:杏鲍菇不同菌株的原生质体的潮霉素敏感性不同,不同菌株菌丝体的潮霉素敏感性相同,相同菌株的原生质体和菌丝体的潮霉素敏感性不同,rs和hph两个基因在杏鲍菇中的共转化率为33%。Objective： The research laid a solid foundation for the improvement of the quality of Pleurotus eryngii （P. eryngii） varities and production of high value products by using P. eryngii as a novel bioreactor. Method： P. eryngii Pel8 and P811 strains were used as materi-Is in this research. The resveratrol synthase gene （rs） and hygromycin phosphotransferase gene （hph） were co - transformed into the pro-toplasts of P. eryngii by PEG-mediated method. Result： The minimal concentration of hygromycin （HmB） which inhibited the growth of Pel 8 protoplasts and P811 protoplasts, respectively, is 201μg./mL and 10μg/mL. Otherwise, the minimal concentration of HmB which inhibi-ted the growth of mycelium from different P. eryngii strains were the same, which is 160μg/mL. 30 out of 90 putative transformants were in-tegrated into both rs and hph by PCR amplification with primes. Conclusion： The sensitivity of protoplasts from different P. eryngii strains are not the samel Otherwise, the minimal concentration of HmB which inhibited the growth of mycelium from different P. eryngii strains were the same. Further more, the minimal concentration of HmB which inhibited the growth of protoplasts and mycelium from the same strain is not the same. The co - transformation efficiency of rs gene and hph gene with P. eryngii was 33%.

关 键 词：杏鲍菇 遗传转化 白藜芦醇合酶基因 潮霉素筛选 

分 类 号：Q785[生物学—分子生物学]