Construction And Sequence Analyzing Of The Recombinant Plasmid Affecting Gene HPV6bL1 Translation By RNA Interfering

Objective Condyloma acuminatum (CA) is a hyperplastic disease caused by Human Papillomavirus (HPV) and locates at external genitals and perianal area HPV infection is widespread in the population and associated with a broad spectrum of human diseases ranging papillomas to anogenital from benign condyloma, common skin warts, laryngeal cancers and cervical cancer. In this study, a recombinant plasmid generating short hairpin RNA was constructed — To construct the recombinant plasmid carrying shRNA to HPV6bL1 and analyze the nucleicacid sequence for further searching new gene therapy method of cauliflower excrescence.Methods (1) Three oligo nucleotides HPV6bL1 cDNA were designed referring to that of GenBank then double-stranded RNA was derived through annealing, finally cloned into pSilencer2.1-U6 neo vector digested by two restricted endoenzymes according to its special orientation. And the recombinant plasmid was transformed into DH5a strain. Finally the plasmid identified by restriction enzyme was used for sequence analysis.Results DNA sequencing showed that the recombinant pSilencer2.1-U6 neo carrying shRNA to HPV6bL1 had been constructed and the aim sequence had been obtained.Conclusion The construction of the recombinant plasmid carrying shRNA to HPV6bL1 lays the basis for the study of its inhibitive effect on cauliflower excrescence. Thus provide material foundation to study the function of the protein and HPV vaccine engineering of CA.