中国现代医学杂志2003，Vol.13Issue(18)：14-18,23,6.
转录激活因子AP-2β基因的克隆、表达及其抗体的制备
CLONING,EXPRESSION OF HUMAN TRANSCRIPTION FACTOR AP-2β GENE AND PREPARATION OF ANTI- AP-2β ANTIBODY
摘要
Abstract
Objective: The aim of this research is to clone,express and purify the AP-2β gene and prepare the antibody.Methods:The AP-2β gene was fused to the glutathion-S-transferase gene in the expression vector pGEX-4T-1.The clones were sequenced to confirm the reading frame. The correct clones were then used to transform E.coli BL21 and the expression of fusion protein was induced by isopropyl-β-D-thiogalaotopyranoside (IPTG). The AP-2β fusion protein was purified with the Glutathione Sepharose-4B.The activity of protein was detected by electrophoretic mobility shift assay (EMSA). Then the protein was used to immunize rabbit in order to produce anti- AP-2β serum.Results:The 78KD fusion protein was obtained and anti- AP-2β antibody was successfully prepared. The protein and the antibody have a good biological activity.Conclusions: The transcription factor AP-2β was successfully expressed and the anti- AP-2β antibody was prepared. This study provides a basis for the functional analysis of AP-2β.关键词
AP-2β基因 GST表达系统 融合蛋白 基因表达 抗AP-2β抗体Key words
The AP-2β Gene/GST Expression System/Fusion Protein/Gene Expression/Anti-AP-2β Antibody分类
生物科学引用本文复制引用
肖顺勇,周建林,钟英丽,胡翔,江铁山,张健..转录激活因子AP-2β基因的克隆、表达及其抗体的制备[J].中国现代医学杂志,2003,13(18):14-18,23,6.基金项目
This research was supported by National Science Foundation of China (No.30025009) and Science Foundation of Hunan Province(No. 300 7045) （No.30025009）
