|
Genital warts caused by HPV - 6 and HPV - 11 type of sexually transmitted diseases , the incidence rate continued to increase in recent years worldwide , in Europe, America and even China is one of the most common viral sexually transmitted diseases . To date, no exact effect , means to cure the disease , and its repeated attacks has become a thorny issue of clinicians , relapse is rooted in the latent infection HPVDNA in the organization copy . The present study envisaged if blocking a key protein of HPV DNA replication of E1 expression , can effectively suppress the replication of HPV DNA , to reduce the level of HPV in the organization powerful through the body and the immune system to clear the virus , and ultimately achieve cure genital warts purposes. Therefore, the proposed hammerhead ribozyme cutting HPV E1 gene mRNA, inhibition of HPV DNA replication of ideas . To this end, we focused on HPV - 6b and HPV - 11 E1 gene homologous sequences to design common to both the the ribozyme - Rz1198 and Rz1282 , and in vitro experiments . Use the self- shear the ribozyme carrier p1.5.1 of a 5'- cis - Rz and 3' - cis - Rz were cloned into the genes of the synthetic Rz1198 Rz1282 , in vitro transcription for two ribozyme ; HPV - 6b and HPV - 11 E1 gene fragment was cloned into plasmid T - vector on the in vitro transcription and in vitro transcription to obtain 32 P - labeled target RNA , respectively ribozyme in vitro cutting experiments . The enzyme kinetic analysis showed that the two ribozymes are efficient, specific cleavage activity of two target RNA , and having a synergistic effect ; in order to increase the cutting performance , both jointly clone in the ribozyme vector p1.5.1 into shotgun type ribozyme -pRE1, and the feasibility of cloning was confirmed by in vitro transcription . This research has successfully cloned two hammerhead common to the HPV - 6bE1 and HPV - 11E1 ribozyme - pR1198 and pR1282 and their associated shotgun-type ribozyme - PRE1 , and successfully carried out in vitro transcription of the ribozyme and a lot of preparation . The two ribozyme enzymatic kinetic results show that the designed the the HPV - 6bE1 and HPV - 11E1 hammerhead ribozyme Rz1198 and Rz1282 common good in vitro cleavage activity , build shotgun-type ribozymes PRE1 is expected to become the inhibition of HPV -6b and HPV - 11 DNA replication of new tools, new ways to open up a gene therapy for the ultimate cure for genital warts .
|