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
em Zh
| WPRIM
| ID: wpr-334797
Biblioteca responsável:
WPRO
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.
Texto completo:
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Base de dados:
WPRIM
Assunto principal:
Fisiologia
/
Plasmídeos
/
Vírion
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Proteínas do Envelope Viral
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Proteínas da Matriz Viral
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Vírus da Leucemia Murina
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Vírus da Síndrome Respiratória e Reprodutiva Suína
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Citometria de Fluxo
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Genética
Idioma:
Zh
Revista:
Chinese Journal of Virology
Ano de publicação:
2008
Tipo de documento:
Article