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[Construction and expression of fusion gene expression plasmid HBV preS2S-rhGM-CSF]
Xiao-lan Guo1, Jian-kang Deng, Dao-yin Zhu
1Department of Clinical Laboratory, Affiliated Hospital of North Sichuan Medical College, Nanchong 637000, China.
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.
