閆培生; 李桂舫; 羅信昌; 周啟
華中農業大學農業部農業微生物重點實驗室; 萊陽農學院應用真菌研究室; 華中農業大學農業部農業微生物重點實驗室; 華中農業大學農業部農業微生物重點實驗室 武漢430070; 萊陽農學院應用真菌研究室; 萊陽265200 ; 萊陽265200 ; 武漢430070 ; 武漢430070
【中文摘要】 利用PEG轉化法將攜有潮霉素抗藥性基因的質粒載體導入香菇原生質體內 ,獲得了抗性轉化子 ,但轉化率較低 ,每微克DNA僅有 0 .5個轉化子。點雜交證實 ,外源基因已整合到香菇染色體上 ;不同轉化子在抗性平板上生長速率不同 ,且轉化子有絲分裂是穩定的。香菇轉基因技術的成功 ,為利用該技術克隆香菇內源基因及啟動子 ,將外源有益目的基因導入香菇體內 ,為培育香菇基因工程菌株提供了可能。
【英文摘要】 The PEG mediated transformation system was used to introduce the plasmid with the hygromycin B resistant gene into the protoplasts of Lentinula edodes. The resistant transformants were obtained at the frequency of 0.5/μg DNA. The dot blot hybridization verified that the foreign drug resistant gene had integrated into the genome of Lentinula edodes. Different transformants showed different growth rate at the same hygromycin contained agar plate, and the true transformants were stable at mitotic stage. This gene transfer system for Lentinula edodes should enable us to realize the objective for cloning the endogenous genes and promoters from Lentinula edodes or introducing foreign beneficial genes,such as the anti fungicide genes and substrate decomposition genes into Lentinula edodes. Therefore, this system will provide a new opportunity for culturing the engineering strains of Lentinula edodes.
【中文關鍵詞】 食用菌; 香菇; 遺傳轉化; 基因工程
【英文關鍵詞】 Edible fungi; Lentinula edodes; Genetic transformation; Gene engineering
【基金】瑞典國際科學基金資助~~
【文獻出處】 食用菌學報,Acta Edulis Fungi,編輯部郵箱,2002年01期 【DOI】CNKI:SUN:SYJB.0.2002-01-001