Cloning and Expression Analysis of FvALT from Fusarium verticillioides
[Objective]Fusarium verticillioides is the main pathogen of maize tassel rot,thus exploring the biological function of FvALT will provide a theoretical basis for the development of novel targeted fungicides.[Method]Physiological and biochemical characteristics and homology of FvALT were analyzed using online tools such as ProtParam,ProtScale,WoLF PSORT and Clustal X.The protein structure was obtained using SOMPA and SWISS-MODEL,and the binding ability of FvALT to substrates was analyzed using AutoDock.The target proteins were then obtained by prokaryotic expression and affinity nickel column chromatography,and the binding ability of FvALT to the substrate was verified by fluorescence titration technique.[Result]FvALT is a hydrophilic protein and localized in the cytoplasm,its protein secondary structure is dominated by α-helix and irregular coiling,and is highly conserved in Fusarium,with a typical PTZ00377 superfamily structural domain.The optimal conditions for the induction of FvALT are 0.4 mol/L IPTG,16℃ for 16 h.FvALT has strong binding effects withα-ketoglutarate,and the binding sites are ARC310,SER149,SER298,SER300 and ASP258.FvALT has a strong binding effect witb α-ketoglutarate,and the binding sites are ARG310,SER149,SER298,SER300,and ASP258.[Conclusion]FvALT has a typical alanine aminotransferase profile and binds specifically to the substrate alpha-ketoglutarate.
万方数据
维普
