Construction and identification of eukaryotic co-expression vector carrying Myod1 and Myog / 中国组织工程研究
Chinese Journal of Tissue Engineering Research
;
(53): 6645-6651, 2013.
Artigo
em Chinês
| WPRIM
| ID: wpr-438530
ABSTRACT
BACKGROUND:
Now it has cooperation and facilitative effete between myogenic regulatory factors through a long time study. So, gene therapy of double genes of Myod1 and Myog can obtain better effect, and can provide a new way for preventing denervated skeletal muscle atrophy.OBJECTIVE:
To construct eukaryotic co-expression vector carrying Myod1 and Myog genes.METHODS:
Ful-length Myod1 gene and Myog gene cDNA were amplified by reverse transcription PCR, and then inserted into pVAX1 vector after digested to establish the recombined Myod1 and Myog eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and then identified with gene sequencing. The in vitro cultured 3T3 cel s were transfected with pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, and the expressions of Myod1 and Myog genes in the 3T3 cel s were detected with western blot assay in order to identify whether the 3T3 cel s could express the target protein correctly. RESULTS ANDCONCLUSION:
The sequencing results showed that the sequence length and base sequence of Myod1 and Myog cDNA in eukaryotic co-expression vector pVAX1-Myod1-IRES2-Myog-IRES2-EGFP were identical with the reported sequences. Myod1 and Myog protein band expressions were detected in 3T3 cel s by western blot after transient transfection. The pVAX1-Myod1-IRES2-Myog-IRES2-EGFP, a eukaryotic co-expression vector of Myod1 gene and Myog gene is successful y constructed.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Tipo de estudo:
Estudo diagnóstico
/
Estudo prognóstico
Idioma:
Chinês
Revista:
Chinese Journal of Tissue Engineering Research
Ano de publicação:
2013
Tipo de documento:
Artigo
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