Pseudotyping of murine leukemia virus particles with porcine reproductive and respiratory syndrome virus M protein-mediated E protein / 病毒学报
Chinese Journal of Virology
;
(6): 345-351, 2008.
Article
in Chinese
| WPRIM
| ID: wpr-334797
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.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Physiology
/
Plasmids
/
Virion
/
Viral Envelope Proteins
/
Viral Matrix Proteins
/
Leukemia Virus, Murine
/
Porcine respiratory and reproductive syndrome virus
/
Flow Cytometry
/
Genetics
Language:
Chinese
Journal:
Chinese Journal of Virology
Year:
2008
Type:
Article
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