SELECTION AND SUBCLONE OF THE TRANSFORMING GENES OF HUMAN PAPILLOMAVIRUS TYPE 16 / 西安交通大学学报(医学版)

ABSTRACT

The transformation of human papillomavirus type 16 (HPV16) may play somc important roles in the oncogenesis of cervical carcinoma Studies showed that the transforming gence mainly located in the E6. E7 open reading frames (ORFs). We digested HPV16 DNA with PstI+EcoRI. Using pUC-19 as the vector, E. coli JM103 strain as the host cell, after two times of directive subelone, plasmid pEP-8 was construeted, confirmed by RE analysis and Southern blot hybridization, the inserted band of pEP-8 was 1.43Kb long contained the intact E6 E7 ORFs, and also a cat-box region, two TATA-boxics and a cell-type specific cnhancer in the upstream swquence. The pEP-8 could be used as the specific probe of HPV-16 E region for identifing virol mRNAs in the HPV infected cells. Meanwhile it was a basework for studying the transformation and transforming proteins of HPV-16.