Screening and identification of interaction proteins of human TIP60? by yeast two- hybrid system
- VernacularTitle:应用酵母双杂交方法筛选与鉴定组蛋白乙酰基转移酶hTIP60?相互作用蛋白
- Author:
Yi HUANG
;
Zhaochun ZENG
- Publication Type:Journal Article
- Keywords:
histone acetyltransferases;
TIP60?;
yeast two-hybrid
- From:Journal of Third Military Medical University
2003;0(14):-
- CountryChina
- Language:Chinese
-
Abstract:
Objective To construct a yeast expression vector containing human TIP60? gene (a spliced form of HIV-1 tat interactive protein,HTATIP,60?103) and screen its interaction proteins by yeast two-hybrid. Methods Human TIP60? gene fragment was amplified by RT-PCR and cloned into pGBKT7 vector. Using TIP60? as bait,the proteins interacting with TIP60? were screened from a human liver cDNA library. The positive clones were analyzed by bioinformatic methods. Results Human TIP60? cDNA was successfully amplified and cloned into the pGBKT7 vector and indentified by double enzyme digestion and DNA sequencing. Using yeast two-hybrid,32 clones were screened from a human liver cDNA library,and 9 positive clones including histone deacetylase 7A (HDAC7A),mouse double minute 2 (DMD2),B-cell CLL/lymphoma 3 (BCL3),endothelin receptor type A (EDNRA),androgen receptor (AR),PHD finger protein 17 (PHF17),Ataxia Telangiectasia Mutated (ATM),CAMP responsive element binding protein 1 (CREB1),orphan steroid receptor BD73 were verified after sequencing. Conclusion The screened interaction proteins with TIP60? by yeast two-hybrid may be involved in the roles of transcriptional regulation of TIP60?.
