A highly efficient polyethylene glycol-mediated transformation method for mushrooms

被引：34
作者：
Li, G
Li, RX
Liu, QY
Wang, Q
Chen, M
Li, BJ [1 ]
机构：
[1] Sun Yat Sen Univ, Minist Educ, Key Lab Gene Engn, Guangzhou 510275, Guangdong, Peoples R China
[2] Zhejiang Gongshang Univ, Dept Biotechnol Engn, Hangzhou, Peoples R China
来源：
FEMS MICROBIOLOGY LETTERS | 2006年 / 256卷 / 02期
关键词：
PEG-mediated transformation; Pleurotus ostreatus; hph; gfp;
D O I：
10.1111/j.1574-6968.2006.00110.x
中图分类号：
Q93 [微生物学];
学科分类号：
071005 ; 100705 ;
摘要：
A highly efficient transformation system mediated by polyethylene glycol was developed for the cultivated mushroom Pleurotus ostreatus. Eighty to 180 integrative and stable-resistant colonies appeared per mu g of DNA per 10(7) viable protoplasts in a transformation experiment with the hygromycin B phosphotransferase gene (hph), which is about 40-1800 times higher than that previously reported in P. ostreatus. One hundred to 150 transformants emitting green fluorescence were observed per mu g of DNA per 10(7) viable protoplasts in a transformation with the green fluorescent protein gene, but green fluorescence disappeared 30 h after transformation, suggesting that the green fluorescent protein gene was only transiently expressed in P. ostreatus. Plasmid pAN7-1 was also transferred into two important cultivated mushrooms, Ganoderma lucidum and Lentinus edodes, and 120-150 and 85-100 transformants per mu g of DNA per 10(7) viable protoplasts were obtained, respectively, which is seven to 38 times and 24-28 times greater than previously reported. These data indicate that this new polyethylene glycol-mediated transformation procedure is highly efficient for mushrooms, and could be a useful tool in mushroom improvement by gene engineering.
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收藏
页码：203 / 208
页数：6
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