【作者】任艷; 黃玉杰; 張新建; 楊合同
【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
【作者機構】山東省科學院生物研究所; 山東省應用微生物重點實驗室
【摘要】 采用根癌農桿菌(Agrobacterium tumefaciens,EHA105菌株)介導法,對平菇(Pleurotus ostreatus)菌絲體遺傳轉化的最佳試驗條件進行研究,結果表明:當平菇菌絲體與甘露醇和山梨醇的混合液預處理20min,且與農桿菌共培養60min時,轉化效率高且用時較少。經PCR和Southern Blotting檢測表明,轉移DNA(transferred DNA,T-DNA)中的潮霉素抗性基因hph已整合到轉化子基因組中。
【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.
【關鍵詞】 平菇; 根癌農桿菌; hph基因; 高滲; 共培養
【Key words】 Pleurotus ostreatus; Agrobacterium tumefaciens; hph gene; hyper-osmosis; co-cultivation
【基金】山東省科技發展計劃項目(編號:2008GG10002025);; 國家高技術研究發展計劃(863計劃)(編號:2006AA10A211)的部分研究內容~~
【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,2010年01期
【DOI】CNKI:SUN:SYJB.0.2010-01-004
【分類號】S646.14