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Vol.29 Suppl.2 时国庆等：雌激素受体ER阝配体结合区的异源表达与纯化 211. 参考文献 [5]Kuiper G C.Enmark E.Pelto-Huikko M.et al.Cloning of a novel receptor expressed in rat prostate and ovary.Proc Natl A- []时国庆，江桂斌.环境化学进展，北京：化学工业出版社， 2005.346 cad Sci USA,1996,93(12):5925 [2]Hall J M,Couse JF,Korach KS.The multifaceted mechanisms [6]Pike A C W,Brzozowski A M.Hubbard R E.et al.Structure of of estradiol and estrogen receptor signaling.J Biol Chem.2001. the ligand-binding domain of oestrogen receptor beta in the pres- 276(40):36869 ence of partial agonist and a full antagonist.EMBO J.1999.18 [3]Shiau A K,Barstad D.Radek J T,et al.Structural characteriza- (17):4608 tion of a subtype-selective ligand reveals a novel mode of estrogen [7]Feng W.Graumann K.Hahn R.et al.Affinity chromatography receptor antagonism.Nat Struct Biol.2002.9(5):359 of human estrogen receptoralpha expressed in Saccharomyces [4]Kuiper GG.Carlsson B.Grandien K.et al.Comparison of the cerevisiae.Combination of heparin and 17beta estradiol affinity ligand binding specificity and transcript tissue distribution of estro- chromatography.JChromatogr A.1999.852(1):161 gen receptors alpha and beta.Endocrinology.1997,138(3):863 Heterologous expression and purification of the ligand binding domain of estrogen receptor B SHI Guoging,GUO Yanfeng,WEI Wei,SONG Qing,ZHANG Huai,ZHONG Guangrong Applied Science School.University of Science and Technology Beijing.Beijing 100083.China ABSTRACI In order to investigate the interaction between environmental chemicals and estrogen receptors, the ligand binding domain of human estrogen receptor(hERLBD)was expressed in Escherichia coli.Based on the optimized expression conditions (IPTG.0.2mmol culture temperature,25C:time of induction. 8h),after purified by affinity chromatography,the production of hERLBD can be reached to 236mg. KEY WORDS estrogen receptor:ligand binding domain:expression:purification参 考 文 献 ［1］ 时国庆江桂斌．环境化学进展．北京：化学工业出版社 2005：346 ［2］ Hall J MCouse J FKorach K S．The multifaceted mechanisms of estradiol and estrogen receptor signaling．J Biol Chem2001 276（40）：36869 ［3］ Shiau A KBarstad DRadek J Tet al．Structural characterization of a subtype-selective ligand reveals a novel mode of estrogen receptor antagonism．Nat Struct Biol20029（5）：359 ［4］ Kuiper G GCarlsson BGrandien Ket al．Comparison of the ligand binding specificity and transcript tissue distribution of estrogen receptors alpha and beta．Endocrinology1997138（3）：863 ［5］ Kuiper G GEnmark EPelto-Huikko Met al．Cloning of a novel receptor expressed in rat prostate and ovary．Proc Natl Acad Sci USA199693（12）：5925 ［6］ Pike A C WBrzozowski A MHubbard R Eet al．Structure of the ligand-binding domain of oestrogen receptor beta in the presence of partial agonist and a full antagonist．EMBO J199918 （17）：4608 ［7］ Feng WGraumann KHahn Ret al．Affinity chromatography of human estrogen receptor-alpha expressed in Saccharomyces cerevisiae．Combination of heparin-and 17beta-estradio-l affinity chromatography．J Chromatogr A1999852（1）：161 Heterologous expression and purification of the ligand binding domain of estrogen receptor β SHI GuoqingGUO Y anfengWEI WeiSONG QingZHA NG HuaiZHONG Guangrong Applied Science SchoolUniversity of Science and Technology BeijingBeijing100083China ABSTRACT In order to investigate the interaction between environmental chemicals and estrogen receptors the ligand binding domain of human estrogen receptor β（hERβ－LBD） was expressed in Escherichia coli．Based on the optimized expression conditions （IPTG0∙2mmol·L －1 ；culture temperature25℃；time of induction 8h）after purified by affinity chromatographythe production of hERβ－LBD can be reached to236mg·L －1． KEY WORDS estrogen receptor；ligand binding domain；expression；purification Vol．29Suppl．2 时国庆等： 雌激素受体 ERβ配体结合区的异源表达与纯化 ·211·