Construction and expression of eukaryotic expression vector of HPV 18E5 and EGFP fusion gene
10.3760/cma.j.issn.1003-9279.2015.01.026
- VernacularTitle:HPV18E5与EGFP融合基因真核载体的构建及表达
- Author:
Fan LI
1
;
Jintao LI
;
Yangjunqi WANG
;
Chuyi LIU
;
Rugang ZHONG
;
Yi ZENG
Author Information
1. 100124,北京工业大学生命科学与生物工程学院环境与病毒肿瘤学北京市重点实验室
- Keywords:
Papillomavirus,human;
Genes;
Rats
- From:
Chinese Journal of Experimental and Clinical Virology
2015;29(1):77-79
- CountryChina
- Language:Chinese
-
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.
