HEV ORF3基因真核表达细胞系HepG2-ORF3的构建
Expression of Swine hepatitis E virus ORF3 gene in HepG2
为了研究猪源戊型肝炎病毒（swHEV）ORF3部分基因的抗原性，尝试通过G418的筛选，建立出稳定表达ORF3蛋白的真核细胞系。试验通过设计引物，扩增 swHEV ORF3部分基因片段，长度360bp。使用pMD18-T载体克隆，双酶切后插入pcDNA3.1真核表达载体。质粒经双酶切鉴定阳性后，转染真核细胞系HepG2，并进行G418筛选。筛选出HepG2-ORF3真核表达细胞系后，通过间接免疫荧光检测其表达情况，同时进行HepG2-ORF3细胞系的形态学和生长曲线的观察。实验最终证实成功建立HepG2-ORF3真核表达细胞系。
o express swine hepatitis E virus (sHEV) ORF3 gene fragment in HepG2, one pair of specific primers were designed. The fragment was cloned into pcDNA3.1 vector and the recombinant plasmid was designated as pcDNA3.1-ORF3. The recombinant plasmid pcDNA3.1-ORF3 was transfected into HepG2 cells for the purpose of establishment of stable expressing pORF3 cell line. Positive clone was screened by G418 with the best screening concentration that depended on the previous experiments. The result showed that the protein expressed was stable, and cell growth curve was shown meanwhile.
梁楚敏、张亮权、纪方晓、王衡、张桂红、梁焕斌、古洪浪
生物科学研究方法、生物科学研究技术基础医学细胞生物学
猪戊型肝炎病毒ORF3 基因真核表达
swine Hepatitis E virusORF3 geneeukaryotic expression
梁楚敏,张亮权,纪方晓,王衡,张桂红,梁焕斌,古洪浪.HEV ORF3基因真核表达细胞系HepG2-ORF3的构建[EB/OL].(2014-03-03)[2025-04-26].http://www.paper.edu.cn/releasepaper/content/201403-19.点此复制
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