Construction of pcDNA3-HERG-G572R expression vector and establishment of a cell line stably expressing HKE-HERG-G572R / 南方医科大学学报
Journal of Southern Medical University
;
(12): 308-311, 2014.
Artículo
en Chino
| WPRIM
| ID: wpr-356931
ABSTRACT
<p><b>OBJECTIVE</b>To construct the pcDNA3-HERG-G572R expression vector and establish a cell line stably expressing HKE-HERG-G572R.</p><p><b>METHODS</b>HERG-G572R mutant fragment was constructed by over-lap extension PCR and validated by DNA sequencing. The HKE-HERG-G572R expression vector was constructed and transfected into HEK293 cells to obtain a cell line stably expressing HKE-HERG-G572R.</p><p><b>RESULTS</b>The pcDNA3-HERG-G572R expression vector was successfully constructed and the cell line stably expressing HKE-HERG-G572R was established. Real-time PCR and Western blotting revealed a 632-fold HKE-HERG-G572R overexpression in the transfected HEK293 cells as compared with that in control HEK293 cells transfected with pcDNA3 (P<0.01).</p><p><b>CONCLUSION</b>The protocol can be used to construct the cell line stably expressing HKE-HERG-G572R to provide a cell model for studying individualized therapy.</p>
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Secuencia de Bases
/
Transfección
/
Expresión Génica
/
Canales de Potasio Éter-A-Go-Go
/
Células HEK293
/
Vectores Genéticos
/
Genética
/
Metabolismo
/
Mutación
Tipo de estudio:
Guía de Práctica Clínica
Límite:
Humanos
Idioma:
Chino
Revista:
Journal of Southern Medical University
Año:
2014
Tipo del documento:
Artículo
Similares
MEDLINE
...
LILACS
LIS