Histone之去乙醯脢1(HDAC1)：免疫化學與生物化學上之鑑定

Abstract

HDAC1是屬於histone去乙醯脢(histone deacetylases)之一成員。而histone去乙醯脢在近年來則是被認為與基因表現的抑制有關。而在本篇論文中，選殖了HDAC1並且純化了重組蛋白以製備抗HDAC1多源性抗體。此外，藉由 pcDNA載體我們成功的在293T細胞中將有帶有FLAG epitope的HDAC1(FLAG–HDAC1)表現出來，而利用免疫螢光染色法觀察到受FLAG–HDAC1轉染後的293T細胞，FLAG－HDAC1於細胞中的位置與內生性之HDAC1是相同的皆是位於細胞核內。而在FLAG–HDAC1之去乙醯化測試中，FLAG－HDAC1的確可將[14C]乙醯基從histone上去除。

HDAC1, a member of histone deacetylases, has been shown to correlate with the repression of the gene expression. In this work, I cloned the HDAC1, purified the recombinant protein and prepared the anti- HDAC 1 antibody. FLAG-tagged HDAC1 was expressed in 293T cells following transfection of HDAC1/pcDNA-tag plasmid. Subcellular localization of the FLAG-tagged HDAC1 was shown to be the same as endogenous HDAC1 (i.e., in the nucleus) by immunofluorescence staining. Furthermore, the FLAG tagged HDAC1 retains the deacetylase activity as demonstrated by the deacetylation of [14C] acetylated histones.