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Optimizing Prokaryotic Expression of a Xylanase Gene from the Apple Pathogen Valsa mali var. mali

並列摘要


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 ß-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.

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