【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(jī)構(gòu)】山東省科學(xué)院生物研究所; 山東省應(yīng)用微生物重點實驗室

【摘要】 采用根癌農(nóng)桿菌(Agrobacterium tumefaciens,EHA105菌株)介導(dǎo)法,對平菇(Pleurotus ostreatus)菌絲體遺傳轉(zhuǎn)化的最佳試驗條件進(jìn)行研究,結(jié)果表明:當(dāng)平菇菌絲體與甘露醇和山梨醇的混合液預(yù)處理20min,且與農(nóng)桿菌共培養(yǎng)60min時,轉(zhuǎn)化效率高且用時較少。經(jīng)PCR和Southern Blotting檢測表明,轉(zhuǎn)移DNA(transferred DNA,T-DNA)中的潮霉素抗性基因hph已整合到轉(zhuǎn)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(guān)鍵詞】 平菇; 根癌農(nóng)桿菌; hph基因; 高滲; 共培養(yǎng)

【Key words】 Pleurotus ostreatus; Agrobacterium tumefaciens; hph gene; hyper-osmosis; co-cultivation

【基金】山東省科技發(fā)展計劃項目(編號:2008GG10002025);; 國家高技術(shù)研究發(fā)展計劃(863計劃)(編號:2006AA10A211)的部分研究內(nèi)容~~

【文獻(xiàn)出處】 食用菌學(xué)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