| 用T-A载体克隆技术构建丙肝病毒C+E1区真核表达质粒pSVL-HCV/C+E1 |
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| 引用本文： | 李益民,周旭,白植生.用T-A载体克隆技术构建丙肝病毒C+E1区真核表达质粒pSVL-HCV/C+E1[J].微生物学免疫学进展,1997(4). |
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| 作者姓名： | 李益民 周旭 白植生 |
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| 作者单位： | 卫生部兰州生物制品研究所 |
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| 摘 要： | 用ＢａｍＨＩ和ＨｉｎｄＩＩ将丙肝病毒Ｃ＋Ｅ１ＤＮＡ片段从其克隆载体ｐＧＥＭ３ｚｆ－ＨＣＶ／Ｃ＋Ｅ１上切下，经Ｔａｑ酶补齐３’末端后插入到载体ｐＳＶＬ－Ｔ中，构建成丙肝病毒Ｃ＋Ｅ１真核表达载体ｐＳＶＬ－ＨＣＶ／Ｃ＋Ｅ１。本实验中重组效率达６４．７％（１１／１７），正向插入为５０％（２／４）。
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| 关 键 词： | 丙肝病毒 T-A载体技术 质粒 |
Construction of the expression vector of hepatitis C virus/C E1--pSVL-HCV/C E1 by T-A cloning technique |
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| Abstract: | The clone plasmid of hepatitis C virus /C+E1 was digested with the restriction enzymes to get the HCV/C+E1 DNA fragment.The single 3'-A was added to C+E1 DNA by Taq DNA polymerase to form C+E1-A DNA fragment.The expression vector of hepatitis C virus/C+E1(pSVL-HCV/C+E1)was constructed by inserting the C+E1-A DNA into pSVL-T eukaryotic vector.In this experiment,the recombinant efficent of C+E1-A and pSVL-T vector was 64.7%(11/17),and the positive-orientation pSVL-HCV/C+E1 reached 50%(2/4).The pSVL-HCV/C+E1 can cause BALB/C mice to produce antibodies against HCV/C. |
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| Keywords: | Hepatitis C T-A cloning technique Plasmid |
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