Production of recombinant vectors for expression of different forms of Kaposi´s Sarcoma Associated Herpesvirus (KSHV) K1 protein

Abstract

Kaposi´s Sarcoma Associated Herpesvirus (KSHV), or Human Herpesvirus 8 (HHV-8), is a gamma Herpesvirus essential for the development of all forms of Kaposi´s sarcoma (KS). The KSHV's life cycle is basically divided into latent and lytic phases, which have distinct viral gene expression profiles. Some important oncogenic products of KSHV are expressed during the lytic phase, including the viral K1 protein. As an effect of interference with intracellular signaling, K1 expression increases the proliferation and survival of KSHV-infected cells. Due to its high level of genetic variability compared to other regions of the viral genome, the K1-encoding ORF (ORF-K1) is commonly evaluated for KSHV genotyping. To date, it is unclear whether different viral genotypes have particular biological effects that might modify the KSHV oncogenicity. The present study aims to contribute to the establishment of an experimental in vitro model for the evaluation of the K1 protein from common KSHV genotypes. Recombinant expression vectors with the ORF-K1 from KSHV genotypes A, B and C will be prepared by genetic cloning. The clones produced will be validated regarding the DNA sequence of the viral ORF-K1, as well as by expression of the K1 protein at the transcriptional level in transfected cells in vitro. As a result, it is expected that recombinant vectors expressing ORF-K1 of KSHV will be successfully produced for use in future studies of the biological properties of the K1 protein from distinct viral genotypes.(AU)