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Promoter trapping in Magnaporthe grisea
Authors:Xiao-hong Liu  Jian-ping Lu  Jiao-yu Wang  Hang Min  Fu-cheng Lin
Institution:(1) Biotechnology Institute, School of Agriculture and Biotechnology, Zhejiang University, Hangzhou, 310029, China;(2) School of Life Sciences, Zhejiang University, Hangzhou, 310029, China
Abstract:Application of promoter trapping based on transformation in Magnaporthe grisea is reported in this paper. Two promoter-trapping vectors, designated as pCBGFP and pEGFPHPH, were constructed and transformed into protoplasts of M. grisea. A library of 1077 transformants resistant to hygromycin B was generated. Of which, 448 transformants were found to express eGFP gene in different structures ofM. grisea. Three transformants grew slowly, 5 transformants decreased in conidiation and 7 transformants reduced in pathogenicity greatly among these 448 transformants. Eleven transformants were checked by genomic southern blot randomly, and 9 of which were single-copy insertions. The promoter trapping technique has been applied successfully in M. grisea and can be used as a tool for functional genomic analysis.
Keywords:Promoter trapping  Green fluorescent protein  Magnaporthe grisea
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