Expression of open reading frame 5 protein of porcine reproductive and respiratory syndrome virus using semliki forest virus expression system.
The ORF5 gene encodes a major envelope glycoprotein (GP5), which is one of the three major proteins of porcine reproductive and respiratory syndrome virus (PRRSV). The GP5 protein has been known to be a 24.5-26 kDa N-glycosylated envelope protein. The GP5 is involved in inducing neutralizing antibodies. For this reason, the GP5 is primary candidate for the PRRSV subunit vaccine. To produce the native form of GP5 in mammalian cells, we have cloned the ORF5 gene from PRRSV CNV-1 into the Semliki Forest virus (SFV)-based expression vector, resulting in recombinant pSFV-ORF5. By the infection with recombinant pSFV-ORF5 to BHK-21 cells, the GP5 expression was confirmed by immunocytochemistry and immunoblotting assay. The recombinant virus particle harboring ORF5 gene was infectious to BHK-21 and MARC-145. The RNA synthesis and expression of GP5 in the infected cell was also confirmed by RT-PCR.
Jung
Hae-Sun
HS
College of Veterinary Medicine, Chungnam National University, Daejeon, 305-764, Korea.
Hwang
In-Wook
IW
Kim
Su-Mi
SM
Kim
Chul-Joong
CJ
Shin
Kwang-Soon
KS
Kim
Hyun-Soo
HS
D000818:Animals; D001483:Base Sequence; D017931:DNA Primers; D005814:Genes, Viral; D010957:Plasmids / Q000235:genetics; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012672:Semliki forest virus / Q000235:genetics*; D013552:Swine; D014759:Viral Envelope Proteins / Q000235:genetics; D014764:Viral Proteins / Q000235:genetics*; D014773:Virology / Q000379:methods
D017931:DNA Primers; D014759:Viral Envelope Proteins; D014764:Viral Proteins; C114789:glycoprotein 5, PRRSV