Cloning of matrix gene of rinderpest virus and expression in eucaryotic cells

2. ABSTRACT The aim of this study was to cloning and in eucaryotic cells to expression of matrix (M) gene of RBOK vaccine strain of rinderpest virus (RPV). For this purpose, total RNA was extracted from Vero cells infected with RBOK vaccine strain of RPV. Matrix DNA amplicons were obtained with reverse transcription polymerase chain reaction (RT-PCR). 1038 bases matrix coding gene region was amplified by using spesific primers to the matrix gene. Then, the M gene was cloned into TOPO XL cloning vector. The presence of M gene was confirmed by PCR screening and enzyme digestion assays. The recombinant plasmid was cut with EcoRL and Sail enzymes to obtain the M gene. Then, the M gene was placed into pCI-neo mammalian expression vector. So, the resulting recombinant plasmid was named as pCI-neo Matrix. Afterwards, the pCI-neo Matrix vector was transiently tranfected into Vero cells. The presence of the Matrix proteins was showed by Western Immunoblotting assay demonstrating that Vero cells express a 38 kDa protein reacting with anti-M antibody. Key words: Rinderpest, matrix gene, cloning, protein expression.