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シナプス終末で働く遺伝子群の共役的発言制御機構の解明

阐明作用于突触末端的基因的协同表达控制机制

基本信息

批准号：
10780364
负责人：
原田真市
金额：
$ 1.47万
依托单位：
Kanazawa 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：
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/en/grant/KAKENHI-PROJECT-10780364/
关键词：
C.elegans unc-18 unc-64 GFP single-strand site-directed mutagenesis

项目摘要

記憶等神経高次機能を制御する神経細胞間情報伝達はシナプスによって担われている。原田はこれまでに、神経伝達物質がシナプス前終末に異常蓄積する変異体の解析から正常なシナプス伝達にはこれらの遺伝子群が神経細胞で共役的に発現することが必須であることを明らかにした。この共役的遺伝子発現の調節機構の解明がシナプス伝達を知る鍵となると考えられる。そこで、(1)シナプス伝達関連遺伝子群の共役的発現制御領域をを明らかにし、(2)神経前終末遺伝子発現の共通性・互換性からシナプス伝達に寄与する因子の遺伝子発現制御ネットワークの構築が本研究の目的である。1.神経特異的な発現制御領域の同定のため線虫C.elegans unc-18およびunc-64ゲノムDNAをクローニングした。PCRで増幅した当該遺伝子上流域DNA断片をGreen Fluorescent Protein(GFP)をレポータ-遺伝子とする発現ベクターに組み込み、トランスジェニック線虫のGFPの発現パターンを観察した。その結果、unc-18遺伝子では上流域-366から-53bpのDNA断片で、またunc-64では上流域-800bpのDNA断片で両者共に神経特異的な発現を示した。2.C.elegansの近種であるC.briggsaeのunc-18上流域をC.elegans unc-18とDNA配列を比較したところC.elegans unc-18上流域-359から-34lbpの19bp範囲で非常に高い相同性を示せす領域を見つけた。しかし、この配列はunc-64中では高い相同性を示す領域は認められなかった。unc-18上流域内の19bpについてsite-directed mutagenesis法により2文字毎に塩基置換を行った発現ベクターを構築し、レポーター活性の局在を指標に神経特異的発現に必須なヌクレオチドを決定した。この結果から19bp内の5'ACX_4SSX_4TTG3'配列がunc-18遺伝子発現に不可欠であることを明らかにした。以上の結果からC.elegans unc-18の神経特異的な遺伝子発現は上記配列に特異的に接合する因子において制御されていることが示唆されたが、unc-64上流域との比較からは共役的発現制御にはさらに別の領域が関与することが推定される。

Memory and other high-level mechanisms are able to control the feelings of the cells. In Harada, in the end of the day, you used to store a lot of money at the end of the day. In the end of the day, you used to store a lot of money at the end of the day. In order to realize the common service of the sub-group of people who work together, it is necessary to make sure that you know how to work together. The children who have served together have found that the institutions have explained that they have reached the level of knowledge and knowledge that they have learned that they are not in the service of each other. In the field of common service, (1) in the field of common service, and (2) in the field of co-operation, (1) in the field of co-operation, and (2) in the field of co-operation, and (2) the purpose of this study is to realize the commonality and interoperability of the sub-group. 1. The current control field of the god is the same as that of C.elegans unc-18, unc-64, DNA, and so on. In the case of PCR, the DNA fragments of the river basin were divided into Green Fluorescent Protein (GFP) fragments and sub-fragments of the watershed. The results showed that the watersheds of the upper watershed-366km-53bp DNA fragments and the watersheds-800bp unc-64 fragments of the unc-18 sub-watersheds showed the same results. The distribution of C.elegans unc-18 DNA on the recent species of 2.C.elegans C. briggsaeae UNC-18 is higher than that of the watershed on the C.elegans unc-18. The 19bp range of the basin-359-34lbp is very high, which indicates that it is very similar in the field of environmental protection. The high homogeneity in the unc-64 indicates that the field is related to each other. In the watershed on the unc-18, the 19bp system site-directed mutagenesis method is based on the text. The text is located in the watershed. The text is located in the watershed. It is necessary to make a decision on the implementation of the information system in the basin. The results show that in 19bp, 5 ACXs, 4 SSXs, 4 TTG3 'are assigned to unc-18 computers, so that you can't owe them to you. The results of the above results show that there is a special joint factor in the C.elegans unc-18 special system, which indicates that the joint factor in the watershed on the unc-64 is more effective than that in the watershed, and that the joint operation in the river basin is in accordance with the presumption of the system.