閆培生; 李桂舫; 羅信昌; 周啟
華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室; 萊陽農(nóng)學(xué)院應(yīng)用真菌研究室; 華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室; 華中農(nóng)業(yè)大學(xué)農(nóng)業(yè)部農(nóng)業(yè)微生物重點實驗室 武漢430070; 萊陽農(nóng)學(xué)院應(yīng)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(zhuǎn)化法將攜有潮霉素抗藥性基因的質(zhì)粒載體導(dǎo)入香菇原生質(zhì)體內(nèi) ,獲得了抗性轉(zhuǎn)化子 ,但轉(zhuǎn)化率較低 ,每微克DNA僅有 0 .5個轉(zhuǎn)化子。點雜交證實 ,外源基因已整合到香菇染色體上 ;不同轉(zhuǎn)化子在抗性平板上生長速率不同 ,且轉(zhuǎn)化子有絲分裂是穩(wěn)定的。香菇轉(zhuǎn)基因技術(shù)的成功 ,為利用該技術(shù)克隆香菇內(nèi)源基因及啟動子 ,將外源有益目的基因?qū)胂愎襟w內(nèi) ,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(guān)鍵詞】 食用菌; 香菇; 遺傳轉(zhuǎn)化; 基因工程
【英文關(guān)鍵詞】 Edible fungi; Lentinula edodes; Genetic transformation; Gene engineering
【基金】瑞典國際科學(xué)基金資助~~

【文獻(xiàn)出處】 食用菌學(xué)報,Acta Edulis Fungi,編輯部郵箱,2002年01期 【DOI】CNKI:SUN:SYJB.0.2002-01-001