Construction and functional identification of eukaryotic expression vector carrying Sprague-Dawley rat MSX-2 gene / 中华整形外科杂志
Chinese Journal of Plastic Surgery
;
(6): 58-62, 2008.
Artículo
en Chino
| WPRIM
| ID: wpr-314159
ABSTRACT
<p><b>OBJECTIVE</b>To construct a high effective eukaryotic expressing plasmid PcDNA 3.1-MSX-2 encoding Sprague-Dawley rat MSX-2 gene for the further study of MSX-2 gene function.</p><p><b>METHODS</b>The full length SD rat MSX-2 gene was amplified by PCR, and the full length DNA was inserted in the PMD1 8-T vector. It was isolated by restriction enzyme digest with BamHI and Xhol, then ligated into the cloning site of the PcDNA3.1 expression plasmid. The positive recombinant was identified by PCR analysis, restriction endonudease analysis and sequence analysis. Expression of RNA and protein was detected by RT-PCR and Western blot analysis in PcDNA3.1-MSX-2 transfected HEK293 cells.</p><p><b>RESULTS</b>Sequence analysis and restriction endonudease analysis of PcDNA3.1-MSX-2 demonstrated that the position and size of MSX-2 cDNA insertion were consistent with the design. RT-PCR and Western blot analysis showed specific expression of mRNA and protein of MSX-2 in the transfected HEK293 cells.</p><p><b>CONCLUSIONS</b>The high effective eukaryotic expression plasmid PcDNA3.1-MSX-2 encoding Sprague-Dawley Rat MSX-2 gene which is related to craniofacial development can be successfully reconstructed. It may serve as the basis for the further study of MSX-2 gene function.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Plásmidos
/
ARN Mensajero
/
Expresión Génica
/
Genes Homeobox
/
Clonación Molecular
/
Análisis de Secuencia de ADN
/
Ratas Sprague-Dawley
/
Proteínas de Homeodominio
/
Vectores Genéticos
/
Genética
Límite:
Animales
Idioma:
Chino
Revista:
Chinese Journal of Plastic Surgery
Año:
2008
Tipo del documento:
Artículo
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