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, 0(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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