Determination of protein protein interactions among protein kinases of amsacta moorei entomopoxvirus (AMEV) and protein kinases with the other proteins of AMEV

Entomopoxviruses are an important group of virus, which infect only insects. These are from poxviruses group which can, as a family, infect both invertebrates and vertebrates, including humans. Protein kinases were known to have roles at virus morphogenesis, host selectivity, the regulation of cell division and apoptosis in some vertebrate poxviruses. In this study, interactions between 2 protein kinases and 230 virus genomic proteins of Amsacta moorei entomopoxvirus have been investigated by yeast-two-hybrid system. The protein kinases genes were cloned in bait and virus genomic genes were cloned in prey vectors. Bait vector was introduced into Saccharomyces cerevisiae AH109. Expression of the bait genes was confirmed by western blot hybridisation analysis. After confirmation, prey vectors were transformed into yeast cells, which carry the bait vectors. Yeast cells contain bait and prey vectors were grown on a selective medium (minimal synthetic defined) to determine the protein-protein interactions between BAIT and PREY proteins. The colonies grown on this medium indicate positive interactions between respective proteins. 5 and 11 interactions were determined among AMV153 and AMV197 protein kinase proteins, respectively. The interacted proteins of AMEV protein kinases and their roles at virus replication will be identified.