| 鸡柔嫩艾美耳球虫EtMIC-2基因真核表达载体的构建及其在Hela细胞中的表达 |
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| 引用本文： | 刘颖丽,李建华,郑君,张西臣.鸡柔嫩艾美耳球虫EtMIC-2基因真核表达载体的构建及其在Hela细胞中的表达[J].中国兽医学报,2011,31(8). |
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| 作者姓名： | 刘颖丽 李建华 郑君 张西臣 |
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| 作者单位： | 吉林大学畜牧兽医学院,吉林长春,130062 |
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| 基金项目： | 吉林省杰出青年计划项目(20070135) |
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| 摘 要： | 根据GenBank发表EtMIC-2序列设计1对特异性引物,通过PCR技术扩增EtMIC-2基因,将该基因与真核表达载体pVAX1连接,构建了重组真核表达载体pVAX1-EtMIC-2,并用脂质体介导法将其转染Hela细胞。然后通过间接免疫荧光（IFA）、SDS-PAGE和Western-blot分析检测EtMIC-2蛋白在转染细胞中的表达情况。经测序和序列分析,成功扩增获得了EtMIC-2基因,并成功构建了重组真核表达载体pVAX1-EtMIC-2,Western-blot和IFA表明EtMIC-2蛋白在Hela细胞中得到表达。该结果为柔嫩艾美耳球虫核酸疫苗的研究奠定基础。
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| 关 键 词： | 柔嫩艾美耳球虫 EtMIC-2基因 真核表达载体 Hela细胞 |
Construction of an eukaryotic expression vector containing EtMIC-2 gene of Eimeria tenella and its expression in Hela cells |
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| LIU Ying-li,LI Jian-hua,ZHENG Jun,ZHANG Xi-chen.Construction of an eukaryotic expression vector containing EtMIC-2 gene of Eimeria tenella and its expression in Hela cells[J].Chinese Journal of Veterinary Science,2011,31(8). |
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| Authors: | LIU Ying-li LI Jian-hua ZHENG Jun ZHANG Xi-chen |
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| Institution: | LIU Ying-li,LI Jian-hua,ZHENG Jun,ZHANG Xi-chen(College of Animal Science and Veterinary Medicine,Jilin University,Changchun 130062,China) |
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| Abstract: | A pair of primers was designed according to the nucleotide sequence of EtMIC-2 gene.The open reading frame(ORF) sequence of EtMIC-2 gene was amplified by PCR,cloned into pVAX1 vector to form the recombinant plasmid pVAX1-EtMIC-2.The pVAX1-EtMIC-2 plasmid was transfected into Hela cells by with lipofection strategy.Recombinant protein expression from the plasmid in Hela cells was confirmed by IFA,SDS-PAGE and Western-blot analysis.The results showed that the EtMIC-2 ORF was amplified correctly by sequences a... |
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| Keywords: | Eimeria tenella EtMIC-2 gene eukaryotic expression vector Hela cell |
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