外源抗药性基因导入香菇体内的研究

Introduction of Foreign Drug-Resistant Gene into Lentinula edodes

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。

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.