食道癌に関与する未知なる遺伝子の同定とその構造及び機能の解明

食管癌未知基因的鉴定及其结构和功能的阐明

• 批准号: 10877198
• 负责人: 橋本 修
• 金额: $ 1.09万
• 依托单位: Keio University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Exploratory Research
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10877198/
• 关键词: 食道癌; ゲノムスキャンニング; RLGS法; クローニング; RLGS; ゲノム解析; 未知遺伝子

食道癌(扁平上皮癌)の遺伝子変化を、ゲノムDNAを2次元電気泳動して解析するRestriction landmark genomic scanning法(RLGS法)を用いて、検討している。食道癌の新鮮手術材料(5症例)の癌部、非癌部組織を採取し、ゲノムDNAを抽出後、RLGS法を施行した。オートラジオグラフィーで得られたprofileをデジタル化し、約1000のスポット化されたDNA断片を癌部、非癌部間で比較した。症例数がまだ少ないものの、癌部のみに出現するスポット変化を2箇所みとめた。これらのスポット変化と,臨床病理学所見との関連性を様々な角度から解析,検討している段階である。スポット変化のうち,1箇所に対しては,既にクローニングを施行済みである。その際、制限酵素NotI切除断端のみを描出し,他の切除断端からなる遺伝子断片を除去するNot I trapper methodを用いている。クローニングは、trapper施行後のprofileから、目的とするスポットを直接打抜き、DNAを抽出した。DNA量の確保のため、プラスミドに挿入し、大腸菌を用いて形質転換を行い、増幅させた。さらにダイデオキシ法にて、塩基配列を決定した。本遺伝子断片と他遺伝子との相同性の検索では、特記すべき相同性は認めなかったものの、本遺伝子断片をプローブとして、さらなる解折が可能と思われた。さらに,本遺伝子断片の塩基配列よりプライマーを作成し,PCRによる脱メチル化の変化を微小病変やパラフィン切片からも検出し得るため,これを計画している。この方法にて,検索症例数を増加させれば,臨床病理学的所見との関連性が明らかになる可能性が考えられる。また,今後はRLGS profi1e数とスポットクローニング数の増加も目標にしたいと考えている。

Two-dimensional swimming analysis of esophageal cancer (flat epithelioma) was performed by two-dimensional Restriction landmark genomic scanning method (RLGS method). The two-dimensional Restriction landmark genomic scanning method was used. The new hand surgery materials for esophageal cancer (5 cases) were extracted from the tissues of the cancer and non-cancer parts. after the DNA was extracted, the RLGS method was performed. We need to know that there is a difference between the cancerous and non-cancerous parts of the profile, which is about 1000 times higher than that of the DNA fragments. The number of cases was less than that of the cancer department, and the cancer department showed that there was a disease in the cancer department. Please analyze the connection and angle seen by the pathology of the bed. If you don't want to change the situation, you will have to pay attention to the situation. The enzyme-limiting NotI was removed from the broken end, and the broken end was depicted. He removed the broken end from the broken end. The fragment was removed. The NotI trapper method was removed. After the trapper is implemented, the profile will be called, the target will be called directly, and the DNA will be pulled out. DNA makes sure that you can make sure that you don't know what you're going to do, and that you can make sure that you don't know what you're going to do, and that you're going to make sure that you can make sure that you don't know what you're going to do. Please tell me that the method and the basis of the column are determined. In this article, we know that the fragments are similar, that they are the same, that they are similar, that they are similar to each other, that they are similar to each other, that they are similar to each other. In this paper, the sub-fragments were arranged in the base line, and the PCR was made. In this case, the minimal disease was removed, and the section was made. The method was used, the number of cases of cords was increased, and what was seen in the pathology of the bed showed that there was a possibility of interconnectedness. In the future, RLGS profi1e will count your numbers and add your eyes to the test.