Construction of the eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 and green fluorescent protein / 南方医科大学学报
Journal of Southern Medical University
;
(12): 433-436, 2009.
Artículo
en Chino
| WPRIM
| ID: wpr-233770
ABSTRACT
<p><b>OBJECTIVE</b>To construct an eukaryotic coexpression vector containing Mycobacterium tuberculosis heat shock protein 70 (mtHSP70) and enhanced green fluorescent protein (EGFP) controlled by cytomegalovirus promoter using pIRES-EGFP vector.</p><p><b>METHODS</b>The mtHSP70 gene fragment was amplified by PCR from pVAX-mtHSP70-HSV2gD using specific primers. The PCR product was cloned into the vector pMD 18-T vector, and the correct clone was selected according to DNA sequence analysis. The interested mtHSP70 gene fragment was subcloned into pCMV-IRES-EGFP vector with XhoI and EcoR I digestion. The recombinant plasmid was transfected into mouse melanoma B16 cell line, and the green fluorescent cells were detected by fluorescence microscopy and mtHSP70 expression was detected by Western blotting.</p><p><b>RESULTS</b>The recombinant plasmid obtained was confirmed by enzyme digestion. The transfected mouse melanoma B16 cells exhibited green fluorescence under fluorescence microscopy and expressed mtHSP70 protein as demonstrated by Western blotting.</p><p><b>CONCLUSION</b>The eukaryotic coexpression vector PCMV-mtHSP70-IRES-EGFP has been established to allow further investigation of the role of mtHSP70 gene in tumor immunotherapy.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes de Fusión
/
Datos de Secuencia Molecular
/
Secuencia de Bases
/
Análisis de Secuencia de ADN
/
Proteínas HSP70 de Choque Térmico
/
Vacunas contra el Cáncer
/
Citomegalovirus
/
Línea Celular Tumoral
/
Proteínas Fluorescentes Verdes
/
Vectores Genéticos
Límite:
Animales
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2009
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS