水稻OsVDAC5真核表达载体的构建及在酵母中的表达研究

Construction of Eukaryotic Expression Vector of OsVDAC5 and Its Expression in Yeast

[目的]构建水稻OsVDAC5的真核表达载体,对其进行真核表达研究。[方法]将Osvdac5基因与酵母表达载体pPIC3.5K连接,通过电转化的方法插入到毕氏酵母GS115中,筛选阳性重组菌株,通过0.5%的甲醇诱导使外源的水稻Osvdac5基因表达,并利用West-ern Blot鉴定表达蛋白的特异性。[结果]成功构建了Osvdac5::pPIC3.5K酵母表达载体,将其整合到酵母GS115基因组后,对其进行诱导表达,产物经Western Blot检测,证实了表达蛋白为OsVDAC5。[结论]该研究结果为进一步研究OsVDAC5蛋白的功能奠定了基础。

[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 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