| 登革2型病毒43株prM基因片段在痘苗中的克隆与表达 |
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| 引用本文： | 孙蕾,杨佩英,秦鄂德,于曼,徐品芳,阎国珍.登革2型病毒43株prM基因片段在痘苗中的克隆与表达[J].生物技术通讯,1998(1). |
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| 作者姓名： | 孙蕾 杨佩英 秦鄂德 于曼 徐品芳 阎国珍 |
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| 作者单位： | 军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所，军事医学科学院微生物流行病研究所 北京 100850，北京 100850，北京 100850，北京 100850，北京 100850，北京 100850 |
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| 摘 要： | 利用RT-PCR技术获得登革2型病毒中国分离株(D_2-43)的prM基因片段。扩增产物经酚:氯仿抽提纯化之后,直接插入pT_7BlueT载体中,经DNA序列分析证明了所扩增片段序列的正确性。构建了痘苗病毒载体PJSA1175/prM,外源基因受痘苗病毒启动子P_(7.5K)的调控。脂质转染(Lipofection)法获得D_2-43株prM蛋白的重组病毒。荧光检测显示,重组痘苗病毒表达的prM蛋白分布于细胞表面。间接ELISA测定其滴度为1:4。
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| 关 键 词： | 登革2型病毒43株 膜蛋白前体(prM) RT-PCR 共转染 基因表达 |
Cloning and expression of prM gene fragment of D2-43 in vaccinia virus |
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| Sun Lei,Yang Peiying,Qin E-de,Yu Man,Xu Pinfang,Yan Guozhen.Cloning and expression of prM gene fragment of D2-43 in vaccinia virus[J].Letters in Biotechnology,1998(1). |
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| Authors: | Sun Lei Yang Peiying Qin E-de Yu Man Xu Pinfang Yan Guozhen |
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| Abstract: | The prM gent fragment of Dengue 2 virus China isolate (D2-43) was obtained by RT-PCR technique. After extrated and purified with phenol: chloroform, the amplified product was directly inserted into pT7-Blue T vector. Nucleotide sequence of the amplified gene fragment was confirmed by DNA sequencing analysis. Then we constructed vaccinia virus recombinant plasmid JSA1175/prM in which the foreign geng was controlled by vaccinia virus promotor P7 5k. Using lipofectin-transfection method, we aquired D2-43 prM recombinant vaccinia virus. Indirect immunosorbent assay showed that the recombinant vaccinia virus expressed prM protein was anchored on cell surface. Result of enzyme-linked immunosorbent test showed that the antigen liter was 1 : 4. |
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| Keywords: | dengue 2 virus-43 prM RT-PCR cotransfection recombinant vaccinia gene expression |
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