J Korean Soc Virol.
1998 Dec;28(4):295-302.
Development of A Monkey Kidney Cell Line Which Expresses Poliovirus Capsid Protein
Abstract
- The RNA genome of poliovirus encodes a long polyprotein precursor and this polyprotein is cleaved proteolytically by viral protease to yield mature proteins. The mature proteins derived from the Pl polyprotein precursor are the component of capsids. To further delineate the process of capsid assembly and encapsidation, in a first attempt, a cell line which expresses the authentic Pl polyprotein was established. CV-1 cells were transfected with the pRCRSVS1P1 plasmid DNA which contains 5'ncr sequences, whole authentic capsid gene of poliovirus and neomycin resistance gene. These cells were treated with G418 for 3 months, and eventually G418 resistant cells were selected and formed colonies. Each colony was picked and grown in the media containing G418. DNA analysis indicated that 1 of 13 neomycin resistant cell lines (R2-18) contains whole poliovirus Pl capsid gene segment which was incorporated into the genome. Immuneprecipitation of cell lysates with sera from rabbit immunized with inactivateded Sabin type 1 particles demonstrated the constitutive expression of the poliovirus Pl capsid protein from R2-18.