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Zhonghua Nan Ke Xue ; 19(1): 15-8, 2013 Jan.
Artículo en Chino | MEDLINE | ID: mdl-23469655

RESUMEN

OBJECTIVE: To construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY. METHODS: PIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot. RESULTS: We successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY. CONCLUSION: The construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.


Asunto(s)
Línea Celular , Proteínas Inhibidoras de STAT Activados/genética , Espermatocitos/citología , Transfección , Animales , Vectores Genéticos , Lentivirus/genética , Masculino , Ratones , Plásmidos
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