Pseudotyping of murine leukemia virus particles with porcine reproductive and respiratory syndrome virus M protein-mediated E protein.
- Author:
Ping-An XIA
1
;
Zhan-Guo DANG
;
Bin ZHOU
;
Pu-Yanz CHEN
;
Bao-An CUI
;
Huang QIU
;
Gao-Feng LU
Author Information
1. The College of Animal Husbandy and Veterinary, Henan Agricultural University, Zhengzhou 450002, China. xpa-88@sohu.com
- Publication Type:Journal Article
- MeSH:
Flow Cytometry;
Leukemia Virus, Murine;
genetics;
Plasmids;
Porcine respiratory and reproductive syndrome virus;
genetics;
Viral Envelope Proteins;
genetics;
physiology;
Viral Matrix Proteins;
genetics;
physiology;
Virion;
genetics
- From:
Chinese Journal of Virology
2008;24(5):345-351
- CountryChina
- Language:Chinese
-
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.
