期刊名称：Advance Journal of Food Science and Technology
印刷版ISSN：2042-4868
电子版ISSN：2042-4876
出版年度：2015
卷号：9
期号：9
页码：701-705
出版社：MAXWELL Science Publication
摘要：Valsa mali var.mali is a causal agent of apple tree canker disease, that causes serious economic losses in China. The lack of understanding of the pathogenic genes and mechanisms of V. mali which poses difficulties to control the disease. Whereas, previous studies have indicated that xylanase produced by V. mali is a key pathogenic factor. In this study, xylanase gene was cloned and obtained recombinant expression vector pET-xylanase, in addition, a prokaryotic expression system for Xylanase protein was established and the expression conditions were optimized. Xylanase protein expressed in Escherichia coli reached a maximum level after induction with 0.5 mmol/L isopropyl &beta-thiogalactopyranoside (IPTG) for 12 h at 30°C. The recombinant protein was confirmed by Western blot analysis using Anti-His mouse monoclonal antibody and the result showed that the induced ~63 kDa band in E. coli was Xylanase protein. The results presented will help to better reveal the xylanase gene functions in the pathogenicity of V. mali.
关键词：Condition optimization ; Prokaryotic expression ; SDS-PAGE ; Western blot ; Xylanase protein ;
