草菇电击法遗传转化的研究  

The genetic transformation of Volvaria volvacea by electroporation

作　　者：吕贝贝[1,2] 于海龙[3] 蒋玮[1,2] 吴潇[1,2] 武国干[1,2] 赵妍[3] 陈蕾[4] 唐雪明[1,2] 

机构地区：[1]上海市农业科学院生物技术研究所,上海201106 [2]上海市农业遗传育种重点实验室,上海201106 [3]上海市农业科学院食用菌研究所,上海201403 [4]南京野生植物综合利用研究院,南京210000

出　　处：《上海农业学报》2017年第3期5-10,共6页Acta Agriculturae Shanghai

基　　金：上海市农委项目农口系统青年成长项目[沪农青字(2016)第1-16号];上海市农委项目[沪农攻科字(2015)第5-6号和第6-1-5号];国家自然科学基金(31500461);上海市农科院人才项目(ZP-17)

摘　　要：建立了草菇电击遗传转化体系,遗传转化的最优条件为:用1.5%溶壁酶,32℃酶解草菇48 h菌龄菌丝体3 h,将20μL 50 ng/μL质粒融入100μL草菇酶解液中,在外加电压1 000 V的条件下电击;电击后的草菇菌丝体涂布于再生培养基中,32℃培养2 d后在含有20μg/mL潮霉素的培养基上进行抗性筛选,分子检测正常生长的单克隆是否为阳性克隆。The genetic transformation system of Volvaria volvacea was established by eletroporation, and the optimal conditions of genetic transformation were as follows：At 32 ℃ the enzymolysis of the mycelia aged 48 h was made with 1.5% lywallzyme for 3 h, and the 100 μL enzymatic hydrolysis solution mixed with 20 μL plasmid at 50 ng/μL was injected electrically under 1 000 voltage;After injected, the protoplast was cultured at 32 ℃ on regeneration medium for 2 d, then transferred to the medium containing 20 μg/mL hygromycin for resistance screening, and lastly the selected normally-growing colonies were verified by molecular identification.

关 键 词：草菇 遗传转化 电击法 潮霉素 

分 类 号：S646.1[农业科学—蔬菜学]