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学科主题: 天然产物有机化学
题名: Over-expression and purification of isotopically labeled recombinant ligand-binding domain of orphan nuclear receptor human B1-binding factor/human liver receptor homologue 1 for NMR studies over-expression; purification; recombinant LBD
其他题名: 同位素标记的人核激素受体hB1F配体结合域的表达和纯化
作者: Chen X(陈翔) ; Tong XT(童孝田) ; Xie YH(谢幼华) ; Wang Y(汪垣) ; Ma JB(麻锦彪) ; Gao DM(高大明) ; Wu HM(吴厚铭) ; Chen HB(陈海宝)
通讯作者: 吴厚铭 ; 陈海宝
刊名: Protein Expr. Purif.
发表日期: 2006-01-01
卷: 45, 期:1, 页:99-106
收录类别: SCI
部门归属: 上海有机化学研究所生命有机化学国家重点实验室; 上海生物化学与细胞生物学研究所分子生物学国家重点实验室
英文摘要: The human hepatitis B virus enhancer II B1 binding factor (hB1F), which regulates the expression of hepatitis B virus genes, is identified as a nuclear receptor. It regulates several liver-specific genes and plays an important role in the bile acid biosynthesis pathway. A significantly optimized protocol has been worked out to prepare N-15 and/or C-13-labeled hB1F ligand-binding domain in minimal medium with high yields for NMR studies. Under the various conditions optimized for the purification of His(6)-hB1F ligand-binding domain, the yield of the purified protein is estimated to be 25-30 mg from 0.5 L of M9 minimal media. Electrospray ionization mass spectrometry data confirm the correctness of the primary sequence. Dynamic light scattering experiment proves that the protein exists as a monomeric form. In addition, the circular dichroism results show that the protein has a well-regulated secondary structure and a high alpha-helical content in ammonium bicarbonate buffer at 20 degrees C and pH 7.4. Finally, uniformly N-15-labeled protein is characterized by a TROSY-HSQC spectrum, and the dispersion of N-15-H-1 cross-peaks in the spectrum indicates the presence of well-ordered and properly folded protein as a monomer. (C) 2005 Elsevier Inc. All rights reserved.
语种: 英语
相关网址: 查看原文
内容类型: 期刊论文
URI标识: http://ir.sioc.ac.cn/handle/331003/17375
Appears in Collections:上海有机化学研究所_期刊论文

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Recommended Citation:
Chen X,Tong XT,Xie YH,et al. Over-expression and purification of isotopically labeled recombinant ligand-binding domain of orphan nuclear receptor human B1-binding factor/human liver receptor homologue 1 for NMR studies over-expression; purification; recombinant LBD[J]. Protein Expr. Purif.,2006,45(1):99-106.
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