Identification of an activator which specifically binds to thyroid hormone receptor b

特异性结合甲状腺激素受体 b 的激活剂的鉴定

• 批准号: 16590902
• 负责人: KANOU Yasuhiko
• 金额: $ 2.18万
• 依托单位: Nagoya University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2004
• 资助国家: 日本
• 起止时间: 2004 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-16590902/
• 关键词: Resistance for thyroid hormone; thyroid hormone receptors; coactivators; Yeast two-hybrid system; cDNA; Cloning

We tried to identified novel proteins which can specifically bind to thyroid hormone receptor (TR) b using yeast two-hybrid system.{Method}Yeast two-hybrid Matchmaker System 3 (Clontech, Palo Alto, CA, USA) was used to screen for novel TR-interacting proteins. A human TR b full length cDNA was cloned into pGBKT7 plasmid vector which contained GAL4 DNA binding domain. We screened a human liver cDNA library (Clontech) using the human TR cDNA as bait in the yeast two hybrid system.{Results}The mating efficiency and number of clones screened corresponded to the values indicated in the manufacture's instructions.1) In the 1st screening we got 131 clones on the plates supplemented with T3 (+T3). The additional screening was yeast clones were streaked both on the +T3 and -T3 plates. Ninety one clones grew in the T3 dependent manner. The plasmids of these yeast clones were purified and the inserts were sequenced. Two unknown sequences were identified ; No. 60 and 93.2) The full length of 2 clones were cloned from human liver total RNA by RT-PCR. Three different mRNAs of the clone No. 60 and a mRNA of No.93 were found in the liver. To determine whether TR physically interact with either the products of clone 60 or 93, the plasmid for GST fusion proteins were constructed.

我们试图用酵母双杂交系统筛选与甲状腺激素受体(TRb)特异性结合的新蛋白。将人TRb全长基因克隆到含有GAL4DNA结合区的pGBKT7载体中。[结果]在酵母双杂交系统中，以人TRc DNA为诱饵，对人肝脏cDNA文库(Clontech)进行筛选.[结果]在第1次筛选中，在添加T3(+T3)的平板上获得131个克隆，其交配效率和克隆数符合生产说明书的要求.额外的筛选是酵母克隆在+T3和-T3平板上都有条纹。91个无性系以T3依赖方式生长。对这些酵母克隆的质粒进行了纯化，并对插入片段进行了测序。通过RT-PCR从人肝总RNA中克隆了2个全长的克隆，分别为第60和93.2号。在肝脏中发现了60号克隆的3个不同的mRNAs和93号克隆的一个mRNAs。为了确定tr是否与克隆60或93的产物发生物理相互作用，构建了GST融合蛋白的表达载体。

项目成果

甲状腺ホルモン受容体遺伝子改変動物.臨床分子内分泌学3-甲状腺・副甲状腺・骨内分泌代謝系-.

甲状腺激素受体转基因动物。临床分子内分泌学3-甲状腺、甲状旁腺和骨内分泌和代谢系统-。

• 发表时间: 2005
• 期刊: 日本臨牀 63(増刊号10)
• 作者: 村田善晴;加納安彦
• 通讯作者: 加納安彦

Spatial and intracellular distribution of the endogenous calcineurin-inhibitory proteins, ZAKI-4, in mouse brain

小鼠大脑中内源性钙调神经磷酸酶抑制蛋白 ZAKI-4 的空间和细胞内分布

• 发表时间: 2004
• 期刊: Acta Histochem.Cytochem. 37・4
• 作者: Peters H;Martini S;Wang Y;Shimizu F;Kawachi H;Kramer S;Neumayer HH.;八幡義人;Hoshino et al.
• 通讯作者: Hoshino et al.

Protective effect of dexamethasone on osmotic-induced demyelination in rats

• DOI: 10.1016/j.expneurol.2004.10.018
• 发表时间: 2005-03-01
• 期刊: EXPERIMENTAL NEUROLOGY
• 影响因子: 5.3
• 作者: Sugimura, Y;Murase, T;Murata, Y
• 通讯作者: Murata, Y

Spatial and intracellular distribution fo the endogenous calcineurin-inhibitory proteins, ZAKI-4, in mouse brain.

小鼠大脑中内源性钙调神经磷酸酶抑制蛋白 ZAKI-4 的空间和细胞内分布。

• 发表时间: 2004
• 期刊: Acta Histochemica et Cytochemica 37
• 作者: Hoshino S;Takagishi Y;Kanou Y;H ayasaka S;Hattori K;Kamabe F;Seo H;Murata Y.
• 通讯作者: Murata Y.

Genomic organization of mouse ZAKI-4 gene that encodes ZAKI-4 α and β isoforms, endogenous calcineurin inhibitors, and changes in the expression of these isoforms by thyroid hormone in adult mouse brain and heart

• DOI: 10.1530/eje.0.1500371
• 发表时间: 2004-03-01
• 期刊: EUROPEAN JOURNAL OF ENDOCRINOLOGY
• 影响因子: 5.8
• 作者: Mizuno, Y;Kanou, Y;Murata, Y
• 通讯作者: Murata, Y