Visualization of DNA binding protein in situ by using double strand DNA fragment, and an attempt of inhibition of the transcription factors' DNA binding by using double strand DNA fragment.

利用双链DNA片段原位观察DNA结合蛋白，并尝试利用双链DNA片段抑制转录因子的DNA结合。

• 批准号: 04557002
• 负责人: TOHYAMA Masaya
• 金额: $ 5.5万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04557002/
• 关键词: transcription factor; DNA binding protein; nuclear transport signal; gene transfer; liposome; histochemistry; nervous system; 転写抑制; 細胞内遺伝子導入; HVJ-リポゾーム; 二本鎖DNAプローブ; HMG1; SV40T抗原; 発現抑制

We investigated following projects1 : Visualization of DNA binding protein in situ by using double strand DNA fragment.2 : An attempt for inhibition of transcription factors'DNA binding by using double strand DNA fragment.For the project3 #1, we have succeeded in establishing the method. We used ^<35>S labeled double strand DNA fragment (Ca.20 base par) which has a AP-1 or CRE motif for the detection of binding protein. The positive signal was observed in cell nucleus.For the project #2, we developed a method which allowed the efficient transportation of DNA fragment into cell nucleus. This was achieved by conjugating nuclear transport signal peptide from SV40 Large T antigen and wrapping by liposome.

我们研究了以下项目1：利用双链DNA片段原位显示DNA结合蛋白。2：利用双链DNA片段抑制转录因子的DNA结合的尝试。我们用S标记的含有AP-1或Cre基序的双链DNA片段(约20个碱基PAR)来检测结合蛋白。在项目2中，我们开发了一种方法，允许DNA片段有效地运输到细胞核中。这是通过将SV40大T抗原的核转运信号肽与脂质体包裹在一起而实现的。

项目成果

H.Kiyama: "Characteristic localization of non-NMDA type glutamate receptor subunits in the rat pituitary gland" Molecular Brain Research. 19. 262-268 (1993)

H.Kiyama：“大鼠垂体中非 NMDA 型谷氨酸受体亚基的特征定位”分子脑研究。

Kiyama H: "Recent Progress in the use of the techniques molecular histochemistry" Jap.J.Thoracic Diseases. 31. 100-107 (1993)

Kiyama H：“分子组织化学技术应用的最新进展”Jap.J.胸部疾病。

Kiyama, H.: "Recent Progress in molecular histochemistry." Acta Anat. Nippon. (in press).

Kiyama, H.：“分子组织化学的最新进展。”

kiyama,H.: "Recent progress in molecular histochemistry." Acta Anat.Nippon. (in press).

kiyama,H.：“分子组织化学的最新进展。”

Koto, K.: "Gene transfer and the expressi on of a foreign gene in vivo in post-mitotic neurons of the adult rat brain using the hemagglutinating virus of the Japan-liposome methd" Mol. Brain Res.25. 359-363 (1994)

Koto，K.：“使用日本脂质体方法的血凝病毒在成年大鼠大脑的有丝分裂后神经元中进行基因转移和体内外源基因的表达”Mol。