2015•Chinese Journal of Clinical HepatologyRequires access
Construction and expression of eukaryotic expression vector of HPV 18E5 and EGFP fusion gene
Fan Li, Jintao Li, Yangjunqi Wang, Chu-Yi Liu, Rugang Zhong
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Abstract
Objective
To construct the eukaryotic expression vector of human papillomavirus type18E5 and EGFP fusion gene, to analyze E5 expression in BALB/c3T3 cell.
Methods
The HPV18E5 and EGFP gene were amplified by polymerase chain reaction, and pSecTag-HPV18 E5-EGFP fusion gene eukaryotic expression vector was constructed by the method of digestion ligation and transformation. The recombinant plasmid was transfected into BALB/c3T3 cells with lipofectamine and the expressed product was detected by RT-PCR, the aim gene expression was observed under fluorescence microscope.
Results
The success in construction of pSecTag- HPV18 E5-EGFP fusion gene eukaryotic expression vector, RT-PCR result showed that HPV18E5 gene could be expressed in BALB/c 3T3 cell, fluorescent light could be observed under fluorescence microscope.
Conclusion
pSecTag-HPV18 E5-EGFP fusion gene eukaryotic expression vector has been successfully constructed, the recombinant plasmid could be expressed in BALB/c3T3 cell.
Key words:
Papillomavirus, human; Genes; Rats