title: [Pseudotyping of murine leukemia virus particles with porcine reproductive and respiratory syndrome virus M protein-mediated E protein].
abstract: For constructing pcDNA-ORF5, pcDNA-ORF5-ORF6, pcDNA-ORF5/6, the ORF5 and ORF6 of porcine reproductive and respiratory syndrome virus (PRRSV) were amplified by RT-PCR, and transiently transfected into 293T cells by calciumphosphate co-precipitation. After 48 h, 293T cells were collected and surveyed by flow cytometry examination. The result indicated that the expression level of the E protein that mediated by the M protein was higher than that of the E protein expressed independently. Then pcDNA-ORF5, pcDNA-ORF5-ORF6, pcDNA-ORF5/6 were respectivly co-transfected into 293T cells with pHIT60 (include MuLV structural genes,namely gag and pol) and pHIT111 (retroviral genome, containing LacZ as a reporter). The supernatants were harvested 48 h post-transfection,and the analysis of the characteristic of the pseudotyping virions was performed by Western blot and infection test. The result indicated that the E proteins were expressed on the virions, and incorporated into the retroviral virions. Infection test were performed on Marc-145 and PAM, all the cells infected were Lac Z positive. These results indicated the pseudotype virions of MuLV-E and MuLV-E/M were infectious, and higher infectivity was achieved by MuLV-E/M.
authors.affiliation: The College of Animal Husbandy and Veterinary, Henan Agricultural University, Zhengzhou 450002, China. xpa-88@sohu.com
authors.forename: Ping-An
authors.initials: PA
authors.lastname: Xia
authors.forename: Zhan-Guo
authors.initials: ZG
authors.lastname: Dang
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.forename: Pu-Yanz
authors.initials: PY
authors.lastname: Chen
authors.forename: Bao-An
authors.initials: BA
authors.lastname: Cui
authors.forename: Huang
authors.initials: H
authors.lastname: Qiu
authors.forename: Gao-Feng
authors.initials: GF
authors.lastname: Lu
mesh_terms: D005434:Flow Cytometry; D009052:Leukemia Virus, Murine / Q000235:genetics*; D010957:Plasmids; D019316:Porcine respiratory and reproductive syndrome virus / Q000235:genetics*; D014759:Viral Envelope Proteins / Q000235:genetics / Q000502:physiology*; D014763:Viral Matrix Proteins / Q000235:genetics / Q000502:physiology*; D014771:Virion / Q000235:genetics*
chemical_list: D014759:Viral Envelope Proteins; D014763:Viral Matrix Proteins