闫培生; 李桂舫; 罗信昌; 周启
华中农业大学农业部农业微生物重点实验室; 莱阳农学院应用真菌研究室; 华中农业大学农业部农业微生物重点实验室; 华中农业大学农业部农业微生物重点实验室 武汉430070; 莱阳农学院应用真菌研究室; 莱阳265200 ; 莱阳265200 ; 武汉430070 ; 武汉430070
【中文摘要】 利用PEG转化法将携有潮霉素抗药性基因的质粒载体导入香菇原生质体内 ,获得了抗性转化子 ,但转化率较低 ,每微克DNA仅有 0 .5个转化子。点杂交证实 ,外源基因已整合到香菇染色体上 ;不同转化子在抗性平板上生长速率不同 ,且转化子有丝分裂是稳定的。香菇转基因技术的成功 ,为利用该技术克隆香菇内源基因及启动子 ,将外源有益目的基因导入香菇体内 ,为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.
【中文关键词】 食用菌; 香菇; 遗传转化; 基因工程
【英文关键词】 Edible fungi; Lentinula edodes; Genetic transformation; Gene engineering
【基金】瑞典国际科学基金资助~~
【文献出处】 食用菌学报,Acta Edulis Fungi,编辑部邮箱,2002年01期 【DOI】CNKI:SUN:SYJB.0.2002-01-001