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Virologica Sinica, 18 (2) : 146-151, 2003
Cloning and Expression in E.coli of Nucleocapsid Protein Gene of Porcine reproductive and respiratory syndrome virus
College of Animal Science and Veterinary Medicine,Huazhong Agricultural University,Wuhan 430070,China
The nucleocapsid protein gene(orf 7)of Porcine reproductive and respiratory syndrome
virus(PRRSV)was isolated from PRRSV genome by RT PCR.The gene was cloned into pMD 1 8一T
vector,the recombinant plasmid pMD1 8N containing orf 7 was sequenced and compared with other
PRRSV isolates.The result shown that the homologies between the cloned orf 7 and America type
PRRSV ATCC VR一2332 reached 100%,the orf 7 was highly conservative sequence in PRRSV genome.
The orf 7 was subcloned into prokaryofic expressing vector pGEX—KG,the recombinant plasmid named
pGEX—KGN was constructed.The pGEX—KGN was used to tran sfo rnl into E colf BL21.Th e results of
SDS—PAGE an d W estem—blot indicated that the nucleocapsid protein gene cloned in downstream of
Glutathione S—tran sferase(GST)was expressed in a high level an d the recombinan t fusion protein,
which was about 41kDa.had immunologicaily reactive activity.This study lay on foundation for th e
development of the diagnosis meth ods in serology fo r PRRSV
  Published online: 15 Apr 2003
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