[Objective] This study aimed to construct the eukaryotic expression vector of OsVDAC5 from rice and investigate its expression in Pichia pastoris.[Method] Osvdac5 gene was ligated to yeast expression vector pPIC3.5K a...[Objective] This study aimed to construct the eukaryotic expression vector of OsVDAC5 from rice and investigate its expression in Pichia pastoris.[Method] Osvdac5 gene was ligated to yeast expression vector pPIC3.5K and inserted into Pichia pastoris GS115 with electroporation to obtain positive recombinant yeast strains. Osvdac5 gene was expressed in yeast after induced with 0.5% methanol, and the specificity of expressed protein was identified by western blotting.[Result] Eukaryotic expression vector Osvdac5∷pPIC3.5K was successfully constructed and transformed into yeast GS115 for the induced expression, and the expressed protein was detected by Western Blotting, which was identified as OsVDAC5.[Conclusion] This study laid the foundation for further investigating the functions of OsVDAC5 protein.展开更多
基金Supported by National Natural Science Foundation of China(31170296)
文摘[Objective] This study aimed to construct the eukaryotic expression vector of OsVDAC5 from rice and investigate its expression in Pichia pastoris.[Method] Osvdac5 gene was ligated to yeast expression vector pPIC3.5K and inserted into Pichia pastoris GS115 with electroporation to obtain positive recombinant yeast strains. Osvdac5 gene was expressed in yeast after induced with 0.5% methanol, and the specificity of expressed protein was identified by western blotting.[Result] Eukaryotic expression vector Osvdac5∷pPIC3.5K was successfully constructed and transformed into yeast GS115 for the induced expression, and the expressed protein was detected by Western Blotting, which was identified as OsVDAC5.[Conclusion] This study laid the foundation for further investigating the functions of OsVDAC5 protein.
