Jin Hua-li, Zhang Fu-chun, Shan Wen-juan, Zhang Ai-lian, Li Yi-jie, Wang Bin
Key Laboratory for Biological resources and genetic engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2004 Sep;20(5):513-6.
To express and identify bovine O type foot and mouth disease virus protein 1 (FMDV VP1) in yeast Pichia pastoris.
FMDV vp1 gene was cloned into secretory Pichia pastoris expression vector-pSuperY. After being linearized with enzyme digestion, the vector was transformed into Pichia pastoris SMD1168H by electroporation. The transformant was screened by zeocin. Expressed proteins in yeast were analyzed by SDS-PAGE and Western blot and then were used to immunize mice.
The results of SDS-PAGE and Western blot demonstrated that the culture supernatant of recombinant yeast contained VP1 protein. The recombinant VP1 protein could elicit similar humoral and cellular immune responses in mice to traditional FMDV killed vaccine.
FMDV VP1 is expressed successfully in yeast Pichia pastoris, which lays the foundation for further FMDV vaccine research.
在毕赤酵母中表达并鉴定牛O型口蹄疫病毒蛋白1（FMDV VP1）。
将口蹄疫病毒vp1基因克隆至分泌型毕赤酵母表达载体-pSuperY。经酶切线性化后，通过电穿孔法将载体转化至毕赤酵母SMD1168H。用博来霉素筛选转化子。对酵母中表达的蛋白进行SDS-PAGE和Western印迹分析，然后用于免疫小鼠。
SDS-PAGE和Western印迹结果表明，重组酵母的培养上清液中含有VP1蛋白。重组VP1蛋白在小鼠中可引发与传统口蹄疫病毒灭活疫苗相似的体液免疫和细胞免疫反应。
FMDV VP1在毕赤酵母中成功表达，为进一步开展口蹄疫病毒疫苗研究奠定了基础。
