摘要
构建融合不同信号肽的O型口蹄疫病毒重组表位蛋白真核表达载体,筛选不同的分泌型信号肽以提高O型口蹄疫病毒重组表位蛋白在中国仓鼠卵巢细胞(Chinese hamster ovary cell,CHO)中的表达并使其分泌到CHO培养基中。通过PCR方法将10种不同的分泌型信号肽与串联表位基因的5′端相连,分别构建了真核表达载体并转染哺乳动物细胞CHO,采用Western-blot检测CHO培养基中重组表位蛋白累积量,研究结果表明,O型口蹄疫病毒重组表位蛋白在CHO中成功分泌表达,不同的分泌型信号肽明显改变O型口蹄疫病毒重组表位蛋白在CHO培养基中的累积SCGB1D1 isoform信号肽介导的O型口蹄疫病毒重组表位蛋白表达量最高。综上所述,合适的分泌型信号肽促进O型口蹄疫病毒重组表位蛋白在CHO中的分泌。
Construction of type O FMDV recombinant epitope prote in eukaryotic expression vector combining different signal peptides,screening different secreted signal peptides to enhance type O FMDV recombinant epitope protein in Chinese hamster ovary cells(CHO).Expressed and secreted it into CHO medium.The 10 different secreted signal peptides were ligated with the 5’end of the tandem epitope gene by PCR,and the eukaryotic expression vector was constructed and transfected into mammalian cell CHO.The recombinant epitope protein in CHO medium was detected by Western-blot.The results showed that the recombinant epitope protein of type O FMDV was successfully secreted and expressed in CHO.Different secreted signal peptides significantly changed the accumulation of recombinant type O FMDV epitope protein in CHO medium,SCGB1D1 isoform signal peptide-mediated expression of the recombinant epitope protein of type O FMDV was the highest.In summary,a suitable secretory signal peptide promotes the secretion of recombinant epitope protein of type O FMDV in CHO.
作者
孙振文
邵军军
常惠芸
SUN Zhen-wen;SHA Jun-jun;CHANG Hui-yun(National Foot-and-Mouth Disease Reference Lab oratory/State Key Laboratory of Veterinary Etiological Biology/Lanzhou Veterinary Research Institute,Chinese A cademy of A gricultural Sciences,Lanzhou 730046,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2020年第3期276-282,共7页
Chinese Veterinary Science
基金
国家重点研发计划项目(2017YFD0500902,2016YFE0204100)。
关键词
表位
蛋白质分泌
口蹄疫病毒
信号肽
epitope
protein secretion
foot-and-mouth disease virus
signal peptide
