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.

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

  • Available online: 05 November 2002
  • 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.

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    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.

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