Related Experiment Videos
[Cloning and expressing of human papillomavirus 16 E7C subgene]
1Department of Biophysics, Institute of Basic Medical Sciences, CAMS and PUMC, Beijing 100005.
Objective:
To develop vaccine against HPV16 is the hotspot in this field, this study was to pave the rudiment for this purpose.
Methods:
E7C subgene (encoding aa39 to aa98) was amplified by PCR, and then cloned into pLNCX plasmid.
Results:
The recombinant pLNCE7C was obtained and identified by gel analysis of the restricted-endonuclease-digested fragments. It was shown that pLNCE7C was expressed in transfected B16 cells. The expression product was identified in the positive cells by Southern blotting and immunohistochemistry method. It was localized mainly in cytoplasm of B16 cells.
Conclusions:
The plasmid pLNCE7C containing subgene of HPV16E7C not only remains the antigenesis of E7 gene but also eliminates the transforming activity of it. Meanwhile the expression of pLNCE7C is quite good. Therefore the plasmid pLNCE7C is suitable to develop DNA vaccine against HPV16.