Construction of a eukaryotic expression vector harboring the small interfering RNA targeting HCMV-IE1 gene and its gene silencing efficiency / 南方医科大学学报
Journal of Southern Medical University
;
(12): 2077-2079, 2010.
Artículo
en Chino
| WPRIM
| ID: wpr-330779
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of RNA interference targeting human cytomegalovirus immediate early gene 1 (HCMV- IE1) on the gene expression in vitro.</p><p><b>METHODS</b>According to the sequence of HCMV-IE1 gene, the small interfering RNA (siRNA) sequences were designed and introduced into the eukaryotic expression vector containing the U6 promoter. After verification by sequence analysis, the recombinant eukaryotic plasmid (pHCMV-IE1i) was transfected into HEL HCMVAD169 cells. The effectiveness of HCMV-IE1 gene silencing was investigated by fluorescence microscopy, flow cytometry and RT-PCR.</p><p><b>RESULTS</b>Sequence analysis confirmed successful construction of the recombinant eukaryotic expression plasmid pHCMV-IE1i. The expression of HCMV-IE1 was effectively suppressed by pHCMV-IE1i transfection in HEL cells as shown by fluorescence microscopy, flow cytometry (P < 0.05) and RT-PCR (P < 0.05).</p><p><b>CONCLUSION</b>The expression of HCMV-IE1 can be effectively suppressed by RNA interference technique in vitro, which provides experimental data for prevention and treatment of HCMV infection.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes
/
Línea Celular
/
Proteínas Inmediatas-Precoces
/
Genes Inmediatos-Precoces
/
ARN Interferente Pequeño
/
Interferencia de ARN
/
Vectores Genéticos
/
Genética
/
Antígenos Virales
Límite:
Humanos
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2010
Tipo del documento:
Artículo
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