O型口蹄疫病毒VP1-2A基因及多表位片段在毕赤酵母中的表达

Expression of Recombinant Pichia pastoris VP1-2A and Multi-epitopes of Type O Foot-and-Mouth Disease Virus

利用毕赤酵母表达系统串联表达O型口蹄疫病毒(FMDV)VP1-2A基因及O型FMDV多表位片段(CTE),将O型FMDVvp1基因和CTE多表位片段克隆到毕赤酵母分泌性表达载体pPIC9K中,构建重组表达载体pPIC9K-VP1-2A-CTE并测序。经SalI线性化后,通过电击转化法将重组质粒导入毕赤酵母GS115中,并对表达产物用SDS-PAGE和Western blot进行分析。重组质粒通过电转进入毕赤酵母后,能表达相对分子量为41.8KD(CTE)和26.5KD(VP1-2A)的蛋白,经Western blot分析,两种蛋白均具有抗原性。在毕赤酵母中成功地表达O型FMDVVP1-2A蛋白和多表位片段(CTE),为研制新型口蹄疫的基因工程疫苗奠定了基础。

To express and identify O-type foot and mouth disease virus protein VP1-2A and CTE epitope fagment in yeast Pichia pastoris.FMDV VP1-2A gene and CTE epitope fagment were cloned into secretory Pichia pastoris expression vector-pPIC9K and sequenced. After being linearized with Sal I enzyme digestion,the vector was transformed into Pichia pastoris GS115 by electroporation. Expressed proteins in yeast were analyzed by SDS-PAGE and Western blot. The results of SDS-PAGE and Western blot demonstrated that the culture supernatant of recombinant yeast contained VP1-2A and CTE protein. FMDV VP1-2A and CTE epitope fagment were expressed successfully in yeast Pichia pastoris,and had good immunogenicity which lays the foundation for further FMDV vaccine research.