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Virologica Sinica, 18 (2) : 146-151, 2003
Contents
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
 Correspondence:
(280.09KB)  
Abstract
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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