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KMID : 0379120110390030235
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Korean Journal of Mycology
2011 Volume.39 No. 3 p.235 ~ p.238
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Agrobacterium-Mediated Transformation of Flammulina velutipes with NaOH Treatment
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Shin Dong-Il
Park Hee-Sung
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Abstract
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Agrobacterium harboring vector pCHBs with hygromycin phosphotransferase(hph) and hepatitis B virus surface antigen(HBsAg)gene was transformed into the mycelial culture of Flammulina velutipes. In particular, mild NaOH solution was treated to the mycelia before Agrobacterium infection step. This was purposed to generate putative surface wounds in the mycelial cell walls. The results showed that hygromycin-resistant(hygr) mycelia could be obtained only from NaOH-treated mycelia but not from intact mycelia. The integration of hygr gene in fungal genome was confirmed by PCR. In addition, a single transgene integration and heterologous protein expression in F. velutipes could be verified by Southern blot hybridization and western blot analysis, respectively. This study demonstrated an efficient tool for the grobacterium-mediated transformation of F. velutipes mycelia.
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KEYWORD
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Agrobacterium, Flammulina velutipes, Mycelium, NaOH, Transformation
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