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国际药用蘑菇期刊
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ISSN 打印: 1521-9437
ISSN 在线: 1940-4344

国际药用蘑菇期刊

DOI: 10.1615/IntJMedMushr.v14.i1.20
pages 21-26

Expression of Hygromycin B Resistance in Oyster Culinary-Medicinal Mushroom, Pleurotus ostreatus (Jacq.:Fr.)P. Kumm. (Higher Basidiomycetes) Using Three Gene Expression Systems

Xiaoya Dong
College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou 450002, China
Ke Zhang
Luoyang Institute of Science and Technology, 90 King City Avenue, Luoyang 471023, China
Yuqian Gao
College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou 450002, China
Yuancheng Qi
Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University, Zhengzhou, P.R. China
Jinwen Shen
College of Life Sciences, Henan Agricultural University, 95 Wenhua Road, Zhengzhou 450002, China
Liyou Qiu
Key Laboratory of Enzyme Engineering of Agricultural Microbiology, Ministry of Agriculture, College of Life Sciences, Henan Agricultural University, Zhengzhou, P.R. China

ABSTRACT

Three hygromycin B phosphotransferase (hph) gene expression systems for culinary-medicinal Oyster mushroom, Pleurotus ostreatus, plasmid pSHC, pAN7-1, and pBHt1 were evaluated through PEG/CaCl2-mediated protoplast transformation. Plasmid pSHC is a newly constructed hph gene expression system, composed of Escherichia coli hph gene, the P. ostreatus sdi promoter, and the CaMV35S terminator. The vector pAN7-1 was commonly used for integrative transformation in filamentous fungi. Plasmid pBHtl is a T-DNA binary vector, usually introduced into fungi by Agrobacterium-mediated transformation. The results showed that plasmids pSHC, pAN7-1, and pBHt1 were all integrated into the host chromosomes and expressed hygromycin B resistance in P. ostreatus. pAN7-1 had the highest transformation efficiency and hph gene expression level, pSHC the second, and pBHt1 the lowest. Growth rates of the transformants on plates containing hygromycin B were in correspondence with their hph gene expression levels. To our knowledge, this is the first report on integrated transformation of plasmid pAN7-1 and pBHt1 in P. ostreatus.


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