title: [Characterization of murine leukemia virus recombinants bearing PRRSV GP5 glycoproteins].
abstract: The highly virulent PRRSV isolate strain HN-1/06 was cultivated on Marc-145. To study the viral entry mechanisms, the GP5 gene of PRRSV isolate was amplified by RT-PCR and cloned into pcDNA3.0 to generate the expressing plasmid pcDNA-GP5. pcDNA-GP5 was transfected into 293T by the calcium phosphate precipitation method. Analysis of flow cytometry confirmed that the GP5 proteins were expressed in surface of the 293T cells. Then 293T cells were transfected with pcDNA-GP5, pHIT60 and pHIT111 plasmids to generate pseudotyping virus. The pseudotyping virus supernatant was harvested 48 hours post-transfection and was detected by Western blotting and infection assay. Western blotting indicated that the GP5 glycoproteins were incorporated into the retroviral pseudotyped virus. Infection assay showed that the pseudotyped virus infected 293T and Mark-145 cell. The pseudotyped virus could be used to further study infectious mechanism of PRRSV.
authors: College of Animal Husbandry and Veterinary, Henan Agricultural University, Zhengzhou 450002, China.
authors: Zhanguo
authors: Z
authors: Dang
authors: Ping'an
authors: P
authors: Xia
authors: Bin
authors: B
authors: Zhou
authors: Yantao
authors: Y
authors: Yin
authors: Jianju
authors: J
authors: Wang
authors: Chunxia
authors: C
authors: Chai
authors: Bao'an
authors: B
authors: Cui
authors: Puyan
authors: P
authors: Chen
mesh_terms: D000818:Animals; D002460:Cell Line; D003001:Cloning, Molecular; D042783:Endothelial Cells / Q000166:cytology / Q000378:metabolism / Q000821:virology; D009052:Leukemia Virus, Murine / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D019316:Porcine respiratory and reproductive syndrome virus / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics; D013552:Swine; D014162:Transfection; D014759:Viral Envelope Proteins / Q000096:biosynthesis / Q000235:genetics*; D014771:Virion / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D014759:Viral Envelope Proteins; C114789:glycoprotein 5, PRRSV