リコビナントgp130を用いたニワトリIL6とLIFのクローニング

使用重组 gp130 克隆鸡 IL6 和 LIF

• 批准号: 12760228
• 负责人: 堀内 浩幸
• 金额: $ 1.41万
• 依托单位: Hiroshima University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Encouragement of Young Scientists (A)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12760228/
• 关键词: ニワトリ; gp130; IL-6; LIF; 抗体; IL6; モノクローナル抗体

本研究では,ニワトリgp130の塩基配列をもとに,リコビナントgp130を作成し,このリコンビナントgp130に対する抗体を作製,利用することにより,gp130のリガンドであるニワトリIL-6とLIFをクローニングすることにある.本年度は昨年度,作製に成功した抗gp130抗体を用いた免疫沈降法とIL-6およびLIFのクローニングを試み,以下の成果を得た.1.作出した抗gp130抗体を用いて免疫沈降法をおこない,リコビナントgp130を特異的に単離できることを確認した.2.ニワトリ由来細胞を用いて1.と同様に免疫沈降をおこない,天然型のgp130を沈降させたが同時に多数のタンパクが沈降することがわかった.3.本研究過程で世界で初めてニワトリLIFのクローニングに成功した.4.同時に,これまでニワトリでは未解析であった血小板由来増殖因子のクローニングにも成功した.5.IL-6の断片クローニングに成功したが,ドイツのグループが先に全長を報告してしまった.6.クローニングしたLIFおよびIL-6のリコンビナント体を作製し,ニワトリ細胞に対して生物活性を持つことを明らかにした.現在,当初の目的通り,これらのリコンビナント体の有効活用系の構築を目指している.

The purpose of this study is to make a study on the basis of gp130, which is made up of gp130, gp130, antibody, and so on. In this study, the antibody was used in this study, and the antibody was used in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did some research in this study. in this study, we did a lot of research in this study. in this study, we did a lot of research in this study. in this study, we did a lot of research in this This year, last year, we successfully used the immunoprecipitation method, IL-6, LIF, LIF, to get the following results. 1. The anti-gp130 antibody was detected by immunoprecipitation method, and the anti-gp130 antibody was confirmed by immunoprecipitation method. 2. The reason for this is that the cell is used in the first place. In the same time, most of the patients were immunized with gp130, while most of them were not immune. 3. In the course of this study, the world was successful in the first half of the year. 4. LIF was successful. At the same time, we did not analyze the origin of progenitor platelets. 5.IL-6 fragments were successfully detected, and the whole report was completed. 6. LIF, IL-6, cell, bioactivity, bioactivity. Now, at that time, there was a general purpose, and there was a flexible use of the system to refer to the target.

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