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アカパンカビDNA修復遺伝子の構造と機能の解析

脉孢菌DNA修复基因的结构和功能分析

基本信息

批准号：
05270202
负责人：
井上弘一
金额：
$ 1.6万
依托单位：
Saitama University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research on Priority Areas
财政年份：
1993
资助国家：
日本
起止时间：
1993 至无数据
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05270202/
关键词：
アカパンカビ除去修復組み換え修復 RAD51ホモログ

项目摘要

本年度は以下の実験を行った。(1)アカパンカビの除去修復遺伝子であるmus-18遺伝子をグルタチオン転移酵素遺伝子に結合し融合タンパク質を大腸菌で発現させた。目的とする大きさのタンパク質を大腸菌から精製し、これを抗原としてウサギに抗体を作らせた。この抗体は野生株の核抽出液と交差反応を示したが、mus-18株のそれとは交差反応を示さなかった。この抗体を用いて現在MUS18タンパク質をアカパンカビから精製している。(2)アカパンカビの組み換え修復遺伝子であるmei-3遺伝子のゲノムおよびcDNAをクローニングし、その塩基配列を決定したところ、これまでに報告のあったmei-3のORFよりさらに上流までORFが伸びていることが明らかとなった。その間に2つのイントロンがあり、新たに70のアミノ酸がN末に加えられた。一方、この新たなORFが正しいことを示すため、いくつかの欠失を持つプラスミドを作成し、これらによるmei-3突然変異株の形質転換を試みた。またノーザンブロット解析によりmei-3遺伝子の転写物の大きさを測定した。これらの結果はいづれも新たに決めたORFが正しいことを示した。NEI3タンパク質は酵母のRAD51遺伝子産物との高い相同性を示した。

This year, the following will be announced as follows. The main results are as follows: (1) in combination with the fusion method, the bacteria were detected by the combination of the enzyme transfer gene and the fusion enzyme. (1) to remove the mus-18 virus, the enzyme transfer enzyme was detected in combination with the fusion enzyme. Objective to measure the concentration of antigens and antibodies in order to detect the essence of bacteria. The wild strain of antibody, nuclear extract solution, and mus-18 strain showed positive reaction, respectively. The antibody was used to detect MUS18 antibodies in the serum. (2) We need to know that we need to mei-3 ORF reports that we need to know that we are going to have an upstream message, that is, we need to know what to do. (2) We need to know that we need to know what to do in the first place. (2) We need to know that we need to know what to do. (2) We need to know that we are going to have a lot of problems. (2) We need to make sure that we know what to do. We need to know what to do. (2) We need to make sure that we can find out what to do. (2) We need to make sure that we know what to do. (2) We need to make sure that we know what to do. We need to know what to do. We need to know what to do. (2) We need to make sure that we know what to do. We need to know I don't know what to do. I'm sorry. I don't know. I don't know. I don't know. On the other hand, the new ORF is showing that it is not valid, and that it is not possible to do so, and that the mei-3 is suddenly in the shape of a strain. I don't know. I don't know what to write. I don't know. I don't know. I don't know what to write. I don't know. The results show that the new ORF is correct and will show you. NEI3 yeast, RAD51 yeast, yeast.