| 猪流行性腹泻病毒S1基因在毕赤酵母中的表达 |
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| 引用本文： | 谭博敏,余希尧,钟泽民,蒋志琼,黄毓茂. 猪流行性腹泻病毒S1基因在毕赤酵母中的表达[J]. 中国畜牧兽医, 2014, 41(6): 64-67 |
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| 作者姓名： | 谭博敏 余希尧 钟泽民 蒋志琼 黄毓茂 |
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| 摘 要： | 为了研制一种能够高效表达猪流行性腹泻病毒S1主要抗原表位基因的新型疫苗，将S1主要抗原表位基因整合到酵母表达载体pPICZαC中，构建了重组质粒pPICZαC-S1，对重组质粒经Sac Ⅰ酶切线性化后通过电转化法转化到毕赤酵母X-33中，经ZeocinTM抗性筛选后得到阳性转化子(pPICZαC-S1)，转化子接种培养基后用甲醇进行诱导，并对诱导结果进行SDS-PAGE分析和Western blotting鉴定。结果表明，纤突蛋白主要抗原表位在毕赤酵母中成功分泌表达，表达的蛋白分子质量为42 ku，且能够与PEDV阳性血清特异性结合。
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Expression of Porcine Epizootic Diarrhea Virus S1 Gene in Pichia pastoris |
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| TAN Bo-min,YU Xi-yao,ZHONG Ze-min,JIANG Zhi-qiong,HUANG Yu-mao. Expression of Porcine Epizootic Diarrhea Virus S1 Gene in Pichia pastoris[J]. China Animal Husbandry & Veterinary Medicine, 2014, 41(6): 64-67 |
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| Authors: | TAN Bo-min YU Xi-yao ZHONG Ze-min JIANG Zhi-qiong HUANG Yu-mao |
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| Abstract: | To develop a new vaccines can efficiently express the porcine epizootic diarrhea virus S1 antigen epitope genes mainly,the recombinant plasmid pPICZαC-S1 was constructed by integrat S1 antigen epitope genes mainly into yeast expression vector pPICZαC and transformed into Pichia pastoris X33 by electroporation after linearized by Sac Ⅰ restriction enzymes. The positive transformants (pPICZαC-S1-X33) were screened by ZeocinTM. The transformants were inoculated into culture medium and induced with methanol. The result was demonstrated by SDS-PAGE and Western blotting, and showed the mainly antigen epitope of spike protein was secreted successfully expression in Pichia pastoris. Expression of the protein molecular weight for 42 ku, and the ability to combine with PEDV positive serum specificity. |
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