Expression and Characterization of Nucleocapsid (N) Gene of R.P.Virus

The book is MVSc research work of the author on recombinant plasmid containing N gene of rinderpest (pBK-CMV-39) of eukaryotic expression of N protein. The fusion protein region along with prokaryotic promoter was removed by Nhel-Spel digestion. The digested plasmid was religated and was used for transforming XL-1 Blue MRF’ cells From the transformed cells the plasmid (pBK-CMV-NS) was purified and characterized by RE digestion. Both the plasmids were used for eukaryotic expression studies.While the cells transfected with pBK-CMV-G9 show expression of a protein of the size 60 kD which corresponds to size of N protein of RPV. The expressed protein reacted with rinderpest polyclonal serum in western blot and ELISA. This indicate that the expressed protein resembles N protein of RPV in immunogenicity. Localization by FAT showed expressed protein was predominantly localized in the cytoplasm of transfected cells. The Number transfected cells were very low.