解放军医学院学报2018，Vol.39Issue(12)：1085-1088,4.DOI:10.3969/j.issn.2095-5227.2018.12.014
人HDAC6基因的原核表达纯化和初步功能检测
Prokaryotic expression and purification of human HDAC6 and preliminary detection of its function
摘要
Abstract
Objective To obtain active purified GST-HDAC6 protein by cloning histone deacetylase 6 (HDAC6) .Methods The coding sequence of HDAC6 was amplified by PCR from human mammary cDNA library, and inserted into the prokaryotic expression vector pGEX-KG.The fusion protein was purified by GST-Sepharose 4B beads, and then identified by SDS-PAGE and Western blot analysis.The biological activity of purified GST-HDAC6 protein was detected by deacetylation experiment.Results From human breast cDNA library, we got a 3645-bp fragment and cloned it into pGEX-KG carrier successfully of which the sequence was in complete accord with target sequence.The target protein expressed by the sequence was with a molecular weight of 170 000 (Mr) from e.coli DH5α.Then, we obtained the highly purified fusion protein GST-HDAC6 after purification.By deacetylation experimental verification, we found the protein showed great enzyme activity.Conclusion HDAC6 is successfully cloned and the active fusion protein is purified, which provide experimental basis for research in relationship between deacetylases and tumor.关键词
HDAC6去乙酰化酶/GST-HDAC6融合蛋白/组蛋白Key words
HDAC6 deacetylases/GST-HDAC6 fusion protein/histone分类
医药卫生引用本文复制引用
陈志达,张鹏飞,郗洪庆,戴广海,卫勃,陈凛..人HDAC6基因的原核表达纯化和初步功能检测[J].解放军医学院学报,2018,39(12):1085-1088,4.基金项目
国家自然科学基金项目(81773135 （）
81572465) （）
