Establish of Molecular-based High-throughput Screening Model for HDAC6 Inhibitors

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	Establish of Molecular-based High-throughput Screening Model for HDAC6 Inhibitors
	Zhang Yue 2; Su Mingbo 2; Zhou Yubo1
刊名	Biotechnology Bulletin
	2012
期号	8 页码:181-188
关键词	HDAC6 HDAC6 Bac-to-Bac Small molecule inhibitors High-throughput screening(HTS)
ISSN号	1002-5464
其他题名	组蛋白去乙酰化酶6(HDAC6)抑制剂分子水平高通量筛选模型的建立
文献子类	Article
英文摘要	It was to establish molecular-based high-throughput screening model for HDAC6 inhibitors.GST-thrombin was subcloned into pFastBac1 vector and then pFastBac1-HDAC6 recombinant plasmid was constructed and transformed into DH10Bac competent cells,generating the recombinant bacmid.Human GST-fusion HDAC6 proteins were expressed in insect Hi5 cells using a baculoviral expression system,and purified using GST sepharose.The deacetylase activity of HDAC6 was assayed with HDAC substrate(Boc-Lys(Ac)-AMC).SAHA was used as the positive control to optimize the activity assay.Results showed the gene segment was confirmed to be the whole coding region of human HDAC6 cDNA by DNA sequencing analysis and the recombinant bacmid was confirmed to have the whole length of HDAC6 cDNA.The GST-HDAC6 fusion protein is expressed in Hi5 insect cells,and purified using GST affinity purification.And the deacetylase activity of HDAC6 was evaluated and confirmed by in vitro assay.Finally,the assay was optimized and the Z-factor is about 0.60,indicating the molecular-based high-throughput screening model for HDAC6 inhibitors was successfully established.
资助项目	国家自然科学基金项目[00000000]
WOS研究方向	Pharmacology & Pharmacy (provided by Clarivate Analytics)
语种	中文
CSCD记录号	CSCD:4604012
内容类型	期刊论文
源URL	[http://119.78.100.183/handle/2S10ELR8/267827]
专题	国家新药筛选中心
通讯作者	Su Mingbo; Zhou Yubo
作者单位	1.Shanghai Institute of Materia Medica, Shanghai 201203, China. 2.East China Normal University, Shanghai 200062, China.;
推荐引用方式 GB/T 7714	Zhang Yue,Su Mingbo,Zhou Yubo. Establish of Molecular-based High-throughput Screening Model for HDAC6 Inhibitors[J]. Biotechnology Bulletin,2012(8):181-188.
APA	Zhang Yue,Su Mingbo,&Zhou Yubo.(2012).Establish of Molecular-based High-throughput Screening Model for HDAC6 Inhibitors.Biotechnology Bulletin(8),181-188.
MLA	Zhang Yue,et al."Establish of Molecular-based High-throughput Screening Model for HDAC6 Inhibitors".Biotechnology Bulletin .8(2012):181-188.