doi:

DOI: 10.3724/SP.J.1145.2013.00986

Chinese Journal of Appplied Environmental Biology (应用与环境生物学报) 2013/19:6 PP.986-989

Cloning and High Expression of HarpinEa USAW004 Gene of Erwinia amylovora USAW004 Strain


Abstract:
From apple shoots infected with fi re blight, we separated and puri fi ed Erwinia amylovora strain USAW004, which caused hypersensitive responses (HR) in non-host plant tobacco leaves. By constructing a USAW004 genomic library and in situ colony hybridization, HarpinEaUSAW004 gene positive clone was screened, and the corresponding fragment was subcloned into the vector pBluescript-II SK(+). An orf of 1 212 bp of HarpinEaUSAW004 gene was determined in the positive clone, with the transcription product HarpinEaUSAW004 consisted of 403 amino acid residues. By cloning HarpinEaUSAW004 gene to the expression vector pET28a(+) under control of the T7 promoter, we successfully constructed an expression plasmid pT7-HrpEaUSAW004. The product HarpinEaUSAW004 was induced by IPTG which expressed in E.coli JM109 with a high effi ciency, also inducing a typical HR in tobacco leaves. Fig 6, Ref 10

Key words:Erwinia amylovora,clone,HarpinEaUSAW004 gene,HarpinEaUSAW004 protein,plant hypersensitive response

ReleaseDate:2015-04-15 08:42:21



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