Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology · 2004
[Construction and expression of fusion gene expression plasmid HBV preS2S-rhGM-CSF].
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Guo Xiao-lan, Deng Jian-kang, Zhu Dao-yin, Tang En-jie
Аннотация
AIM: To construct GM-CSF and preS2 fusion gene expression plasmid to enhance the immunogenicity of hepatitis B DNA vaccine.
METHODS: HBV preS2S gene(846 bp) and rhGM-CSF gene(384 bp) were amplified by PCR, respectively. The eukaryotic expression plasmid pcDNA3.1-S2S-rhGM-CSF was constructed by means of T-A clone and directional gene cloning techniques, and then the recombinant plasmid was expressed in HepG2 cells.
RESULTS: Restriction enzyme digestion analysis, PCR amplification and/or DNA sequencing proved that the recombinant plasmid was constructed successfully. The transcription of target gene was confirmed by RT-PCR. The fusion protein expressed in HepG2 cells could reacted to the monoclonal antibodies (mAbs) against HBsAg, preS2 and GM-CSF, respectively.
CONCLUSION: The successful construction and expression of pcDNA3.1-S2S-rhGM-CSF lay the foundation for further study of HBV DNA vaccine.
METHODS: HBV preS2S gene(846 bp) and rhGM-CSF gene(384 bp) were amplified by PCR, respectively. The eukaryotic expression plasmid pcDNA3.1-S2S-rhGM-CSF was constructed by means of T-A clone and directional gene cloning techniques, and then the recombinant plasmid was expressed in HepG2 cells.
RESULTS: Restriction enzyme digestion analysis, PCR amplification and/or DNA sequencing proved that the recombinant plasmid was constructed successfully. The transcription of target gene was confirmed by RT-PCR. The fusion protein expressed in HepG2 cells could reacted to the monoclonal antibodies (mAbs) against HBsAg, preS2 and GM-CSF, respectively.
CONCLUSION: The successful construction and expression of pcDNA3.1-S2S-rhGM-CSF lay the foundation for further study of HBV DNA vaccine.
Предметные рубрики MeSH
Antibodies, MonoclonalBase SequenceCell Line, TumorGene Expression Regulation, ViralGenes, ViralGranulocyte-Macrophage Colony-Stimulating FactorHepatitis B Surface AntigensHepatitis B virusHepatoblastomaHumansLiver NeoplasmsMolecular Sequence DataPlasmidsProtein PrecursorsRecombinant Fusion ProteinsTransfection