The Transfection And Expression Of Humanpapillovirus 16E7 Gene In Human Laryngeal Carcinoma HEp-2 Cell Line

[Objective] To clone HPV16E7-HSP70 fusion gene and get the fusion protein. To study the effect of humanpapillovirus 16E7 gene to HEp-2 cell line. [Methods] To construct and identify a prokaryotic expression plasmid encoding HPV16E7-HSP70 fusion gene . The plasmid pET28a-HPV16E7-HSP70 was transformed in E. coli BL21 and induced by IPTG. To express , purify and renature HPV16E7-HSP70 fusion protein in E. coli. To construct plasmid pcDNA3.1/myc-His-HPV16E7. and transfected plasmid vector of pcDNA3.1/myc-His-HPV16E7 into the laryngeal carcinoma cells by electroporation.The expression of HPV16E7 was tested by Western blot. [Results] The fusion gene can express successfully in E. coli. HPV16E7 protein was attained from the transfected HEp-2 cells by Western blot. The transfected cells proliferated actively and S phase of cell cycle was prolonged. [Conclusions] The HPV16E7-HSP70 fusion protein could be obtained. Recombinant pcDNA3.1/myc-His-HPV16E7 was transfected into human laryngeal carcinoma cell line successfully by electroporation. HPV16E7 could increase the tumorigenesis of HEp-2 cells.