Construction and identification of a mouse spermatocyte-derived cell line with a stable expression of PIAS-NY.
- Author:
Ying ZHENG
1
;
Hai-Yan WANG
;
Lu-Ping ZHANG
;
Hong-Ya SUN
;
Hong LIANG
;
Xiao-Qing JIA
;
Yan-Qiu HU
;
Yong-Ze ZHU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Cell Line; Genetic Vectors; Lentivirus; genetics; Male; Mice; Plasmids; Protein Inhibitors of Activated STAT; genetics; Spermatocytes; cytology; Transfection
- From: National Journal of Andrology 2013;19(1):15-18
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo 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.
METHODSPIAS-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.
RESULTSWe 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.
CONCLUSIONThe 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.
