CHEN Hongmei BAI Xue—fan, PAN Lei, LI Guang—yu, WEI San—hua, HUANG Chang—xing and Construction and Expression of Eukaryotic Expression Vector Bearing Fusion Gene Of HBV PreS2+S Gene and IFN— Gene[J]. Virologica Sinica, 2002, 17(4): 312-314.
Citation: CHEN Hongmei BAI Xue—fan, PAN Lei, LI Guang—yu, WEI San—hua, HUANG Chang—xing, . Construction and Expression of Eukaryotic Expression Vector Bearing Fusion Gene Of HBV PreS2+S Gene and IFN— Gene .VIROLOGICA SINICA, 2002, 17(4) : 312-314.

HBV PreS2+S/IFN.I】【融合基因真核表达载体的构建及其表达

  • 构建含HBVPrdS2+S和IFN—a融合基因的真核表达载体pcDNA3.1.S2s/IFN.a并在真核细胞中进行表达。 应用重叠延伸剪切技术(splicing by overlapping extension,简称SOE)经两次PCR获得嵌合基因片段S2S/IFN 回 收后直接克隆到pcDNA3.1 V5/His TOPO TA克隆载体,得到真核重组载体pcDNA3.1.S2s/IFN—a。然后用脂质 体法转染vem E6细胞。对重组载体进行了限制性酶切及PCR鉴定,证明连接正确;经间接免疫荧光检测证实该 重组载体能在真核细胞中表达插入的外源性基因编码的融合蛋白。真核表达载体pcDNA3.1.S2s/IFN—a的成功 构建及在vem E6细胞中的有效表达,为进一步探讨HBV感染的特异性免疫治疗提供了实验依据。

Construction and Expression of Eukaryotic Expression Vector Bearing Fusion Gene Of HBV PreS2+S Gene and IFN— Gene

  • To construct a recombinant eukaryotic expression vector bearing fusion gene Of Hepatitis B viruS(HBV)S2+S gene and IFN-a gene,technique of splicing by overlapping extension and two times PCR were used.Fusion gene fragment was obtained and directly cloned into pcDNA3.1 V5/His TOPO TA cloning vactor to get recombinant eukaryotic expression vector pcDNA3.1$2S/IFN—a. Then the recombinant vector was transferred into Vero E6 cells using LipofectAM INE.The recombi— nant vector was constructed and correctly checked by digestion with restriction enzymes and poly— merase chain reaction.The vector bearing fusion gene could be expresed in eukaryotic cells detected by indirect immunofluotescence technique.The relative efficient expression of the fusion gene in Vero E6 cells might provide an experimental basis for specific immunotherapy for HBV infection.

  • 加载中
  • 加载中

Article Metrics

Article views(4232) PDF downloads(829) Cited by(0)

Related
Proportional views
    通讯作者: 陈斌, bchen63@163.com
    • 1. 

      沈阳化工大学材料科学与工程学院 沈阳 110142

    1. 本站搜索
    2. 百度学术搜索
    3. 万方数据库搜索
    4. CNKI搜索

    Construction and Expression of Eukaryotic Expression Vector Bearing Fusion Gene Of HBV PreS2+S Gene and IFN— Gene

    • 1. Department of Infecticious Diseases,Tangdu Hospital,Fourth Military Medical University,Xi an 710038,China

    Abstract: To construct a recombinant eukaryotic expression vector bearing fusion gene Of Hepatitis B viruS(HBV)S2+S gene and IFN-a gene,technique of splicing by overlapping extension and two times PCR were used.Fusion gene fragment was obtained and directly cloned into pcDNA3.1 V5/His TOPO TA cloning vactor to get recombinant eukaryotic expression vector pcDNA3.1$2S/IFN—a. Then the recombinant vector was transferred into Vero E6 cells using LipofectAM INE.The recombi— nant vector was constructed and correctly checked by digestion with restriction enzymes and poly— merase chain reaction.The vector bearing fusion gene could be expresed in eukaryotic cells detected by indirect immunofluotescence technique.The relative efficient expression of the fusion gene in Vero E6 cells might provide an experimental basis for specific immunotherapy for HBV infection.

    Relative (20)

    目录

    /

    DownLoad:  Full-Size Img  PowerPoint
    Return
    Return