Screening PIAS2-interacting proteins in the mouse using the yeast two-hybrid system.
- Author:
Ying ZHENG
1
;
Xiao-Qin JIA
;
Lu-Ping ZHANG
;
Hai-Yan WANG
;
Yan-Qiu HU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Base Sequence; Gene Library; Male; Mice; Protein Binding; Protein Inhibitors of Activated STAT; genetics; Protein Interaction Domains and Motifs; Spermatogenesis; genetics; Spermatogonia; metabolism; Two-Hybrid System Techniques
- From: National Journal of Andrology 2011;17(9):775-780
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo screen and identify PIAS2-interacting proteins from the mouse spermatogonial cDNA library using the yeast two-hybrid system, and to investigate the action mechanism of PIAS2 in spermatogenesis.
METHODSWith pGBKT7-PIAS2 as a bait plasmid, the positive clones interacting with pGBKT7-PIAS2 were screened from the mouse spermatogonial cDNA library, the inserted fragments were sequenced and underwent bioinformatic analysis, and their interaction was verified using the yeast two-hybrid system.
RESULTSThrough screening, sequencing, homology analysis and yeast two-hybrid verification, we obtained 8 different candidate proteins interacting with PIAS2, including Cyfip2, Psmb3, Nmel, nischarin, Ints10, Nsun5, Gnb211 and Ndufaf3.
CONCLUSIONEight different genes were successfully obtained using the yeast two-hybrid system, and their encoding proteins interacted with PIAS2, which might be related with male fertility regulation. Our findings have offered some new clues to the action mechanism of PIAS2 in spermatogenesis.
