Chenyang Jiang, Xinjing Liu, Qingzhi Wang, Ting Yang, Zhiqiang Han, Yuming Xu*
Department of Neurology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, China. firstname.lastname@example.org
Abstract: The element of oriS as a necessary origin sequence for Herpes Simplex Virus type 1derived amplicon vector replicating consists of 45 bp oriS core region and flanking sequences. The flanking sequences of oriS located between gene US12/US1 and RS1 in HSV-1 genome were previously demonstrated to enhance oriS-dependent DNA replication, but the direct evidence of different oriS-flanking sequences in the replication of amplicon virions is still lacked. In this study, we performed a functional analysis of OriS-flanking sequences in replication of HSV-1 based amplicon virions by comparing the yields of virions packaged from amplicon vectors with various deletions of oriS-flanking sequences. The results showed that yields of amplicon virions were significantly reduced by deleting a AgeI/RsrII digested 181bp- fragment between the locus of 333bp-152bp upstream the oriS core region and a NcoI/StyI digested 314bp- fragment between the locus of 23bp-337bp downstream the oriS core region. Furthermore, deletion of a 87bp NcoI/SphI fragment at the locus of 250bp-337bp downstream oriS core region led to a dramatically decrease. Our study further demonstrated that oriS-flanking sequences played an important enhancement role in the amplicon vector replication evidenced at virion level.
[Chenyang Jiang, Xinjing Liu, Qingzhi Wang, Ting Yang, Zhiqiang Han, Yuming Xu. Functional Analysis of OriS-flanking Sequences in Replication of HSV-1 based Amplicon Virions. Life Sci J 2013:10(1):3132-3139]. (ISSN: 1097-8135). http://www.lifesciencesite.com.
Keywords: Functional Analysis; OriS-flanking Sequences; Replication; HSV-1; Virion.