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細胞性粘菌を用いた新規動物型細胞質分裂関連因子の同定と機能解析

利用盘基网柄菌鉴定新型动物型胞质分裂相关因子并进行功能分析

基本信息

批准号：
08J08097
负责人：
島田奈央
金额：
$ 1.54万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for JSPS Fellows
财政年份：
2008
资助国家：
日本
起止时间：
2008 至 2010
项目状态：
已结题

项目摘要

前年度、FACS Caliburを使用し1核当たりに含まれるDNA量を測定した結果、sunB遺伝子破壊株では野生株の約2倍のDNA量を持っていることがわかった。本年度はpropidium iodideにて染色体を検出、顕微鏡下で染色体数を計測した結果、sunB遺伝子破壊株の核では、野生株の2倍である12本の染色体を持つ細胞が多く観察された。また、定量PCRにより各染色体に位置する代表的な遺伝子の1細胞あたりのコピー数を調べた結果もsunB遺伝子破壊株ではほぼ2倍を示した。以上の結果からsunB遺伝子破壊株は2セットの染色体を持つ倍数体である可能性が示された。前年度までにsunB遺伝子破壊株は中心体数に異常がみられたため、その形状の異常についてGFPタグγチューブリンを用いて調べた。その結果、sunB遺伝子破壊株では中心体の面積が野生株よりも約1.5倍大きく、細長い異常な形状であることが明らかになった。これらの結果からSunBは、核分裂と中心体の成長・形状維持に関与していると考えられる。一方、細胞分裂中期のSunBの局在から、染色体の分配に機能するキネトコア複合体との関係性が考えられた。セントロメアのマーカーとしてGFPタグをしたヒストンH3バリアントの局在をsunB遺伝子破壊株で調べた結果、間期の核においても核内に散在するという異所局在が確認された。セントロメアDNA配列の染色体上の転座はみられなかったことから、SunBはタンパク質レベルでの正常なセントロメア構造維持に関わり、この異常が上記の染色体数の異常を引き起こしていると考えられた。

In the previous year, FACS Calibur tested the results by using the first nuclear test, and the results showed that the amount of DNA in the wild strain was about 2 times higher than that in the wild strain, and the amount of DNA in the wild strain was about 2 times higher than that in the wild strain. In this year, propidium iodide chromosomes were isolated, the results of chromosome number calculation under microanalysis, the nuclei of sunB spores were broken, and the chromosomes of wild plants were twice as much as those of wild plants. Quantitative and quantitative PCR were used to determine the number of cells represented by the location of each chromosome. The results showed that the number of broken plants was 2 times higher than that of sunB. The above results showed that the possibility of chromosome retention of sunB spores in the chromosome of the second strain showed significant difference. In the previous year, the number of sunB plants was broken, the number of centrosome, the shape, the shape, the GFP, the γ-body, the body number, the shape, the size, the shape, the shape and the shape of the plant were used. The results showed that the size of the wild plant was about 1.5 times larger than that of the wild plant, and the shape of the wild plant was about 1.5 times larger than that of the wild plant. The shape of the wild plant was about 1.5 times larger than that of the wild plant. The results show that SunB, mitotic centrosome growth and shape maintenance are very important. On the other hand, in the metaphase of cell division, the SunB is in the cell, and the chromosome distribution mechanism is able to detect the sex of the complex. Do you want to know if you want to know if you have a GFP problem? you have to know that the H3 virus has been broken in the sunB and that it has been scattered in the core of the plant. The number of chromosomes on the chromosomes is arranged in an array called DNA. The number of chromosomes on the chromosomes is not affected by the number of chromosomes on the chromosomes. In this way, there is an increase in the number of chromosomes on the chromosomes. The results show that there is an increase in the number of chromosomes on the chromosome.