【作者】任艳; 黄玉杰; 张新建; 杨合同
【Author】REN Yan; HUANG Yujie; ZHANG Xinjian; YANG Hetong Shandong Key Laboratory of Applied Microbiology; Institute of Biology; Shandong Academy of Sciences; Jinan; Shandong 250014; China
【作者机构】山东省科学院生物研究所; 山东省应用微生物重点实验室
【摘要】 采用根癌农杆菌(Agrobacterium tumefaciens,EHA105菌株)介导法,对平菇(Pleurotus ostreatus)菌丝体遗传转化的最佳试验条件进行研究,结果表明:当平菇菌丝体与甘露醇和山梨醇的混合液预处理20min,且与农杆菌共培养60min时,转化效率高且用时较少。经PCR和Southern Blotting检测表明,转移DNA(transferred DNA,T-DNA)中的潮霉素抗性基因hph已整合到转化子基因组中。
【Abstract】 Agrobacterium tumefaciens-mediated transformation of Pleurotus ostreatus was optimal when fungal mycelia were exposed to hyper-osmosis treatment in a mannitol-sorbitol solution and co-cultivation with A. tumefaciens for 20 min and 60 min,respectively. PCR and Southern blotting confirmed that the hygromycin resistance gene (hph) had been integrated into the genomes of P. ostreatus transformants.
【关键词】 平菇; 根癌农杆菌; hph基因; 高渗; 共培养
【Key words】 Pleurotus ostreatus; Agrobacterium tumefaciens; hph gene; hyper-osmosis; co-cultivation
【基金】山东省科技发展计划项目(编号:2008GG10002025);; 国家高技术研究发展计划(863计划)(编号:2006AA10A211)的部分研究内容~~
【文献出处】 食用菌学报,Acta Edulis Fungi,编辑部邮箱,2010年01期
【DOI】CNKI:SUN:SYJB.0.2010-01-004
【分类号】S646.14
