Studies of complicated systems of growth using GH transgenic Amago

使用 GH 转基因 Amago 研究复杂的生长系统

基本信息

批准号：
14360104
负责人：
MORI Tsukasa
金额：
$ 9.41万
依托单位：
Nihon University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2002
资助国家：
日本
起止时间：
2002 至 2004
项目状态：
已结题

项目摘要

(1)Proteins analysis extracted from intestine, liver and serum using DIGEProteins were extracted from intestine, liver and serum of GH transgenic Amago, and these proteins were performed proteins analysis using DIGE. About 2000 proteins were separated by two-dimensional gel electro-phoresisis, and some of major proteins were extracted from the gel and performed MS/MS-analysis. Most of these were unknown proteins, however, amount of digestive enzymes and G-proteins were significantly greater in transgenic intestine than that of control. Analysis of liver tissue also showed vast differences among transgenic and control. There were proteins having similar to parasites amino sequences in transgenic, and these were about four times grater than control. These proteins were detected only all transgenic used for the experiment.(2)Analysis gene expression by microarrayWe have performed subtractive hybridization using liver tissue from GH transgenic and control, and made microarray plates using these subtracted DNA. Analysis of microarray was performed using RNA extracted from both F2 and F3 of immature and mature GH transgenic. The results of these experiments showed drastic decrease of natural immune system in immature young fish. However, this tendency was getting installed when they were matured. Further, some sex related gene expression was observed in immature transgenic, and these tendency were also showed both F2 and F3.(3)Screening of pituitary genes by subtractionThe completed length of cDNA library was obtained using pituitary of immature GH transgenic fish (F2), and also subtractive hybridization was performed using immature GH transgenic. Each 1000 clones of subtracted genes from both higher expression and lower expression were screened, and these have been sequenced.

(1)使用DIGE从肠、肝脏和血清中提取的蛋白质分析从GH转基因Amago的肠、肝脏和血清中提取蛋白质，并使用DIGE对这些蛋白质进行蛋白质分析。通过二维凝胶电泳分离了大约 2000 种蛋白质，并从凝胶中提取了一些主要蛋白质并进行了 MS/MS 分析。其中大部分是未知蛋白质，然而，转基因肠道中消化酶和G蛋白的量显着高于对照。肝组织的分析也显示转基因和对照之间存在巨大差异。转基因中存在与寄生虫氨基酸序列相似的蛋白质，并且这些蛋白质比对照多约四倍。这些蛋白质仅在用于实验的所有转基因中被检测到。(2)通过微阵列分析基因表达我们使用来自GH转基因和对照的肝组织进行了消减杂交，并使用这些消减的DNA制作了微阵列板。使用从未成熟和成熟GH转基因的F2和F3中提取的RNA进行微阵列分析。这些实验的结果表明，未成熟的幼鱼的自然免疫系统急剧下降。然而，当他们成熟时，这种倾向就开始显现。此外，在未成熟转基因鱼中观察到了一些性别相关基因的表达，并且这种趋势在F2和F3中也有表现。 (3)垂体基因的差减筛选利用未成熟GH转基因鱼(F2)的垂体获得cDNA文库全长。，并且还使用未成熟的GH转基因进行消减杂交。对从较高表达和较低表达中减去的基因的每 1000 个克隆进行筛选，并对这些基因进行测序。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Localization of Ghrelin-Producing cells in the stomach of the rainbowtrout(Oncorhynchus mykiss)

虹鳟鱼 (Oncorhynchus mykiss) 胃中生长素释放肽产生细胞的定位

DOI：
发表时间：
2004
期刊：
Zoological Science 21
影响因子：
0
作者：
Ichiro Sakata;Tsukasa Mori;Hiroyuki Kaiya;Mami Yamzki;Kenji Kagwa;Kinji Inoue;Takafumi Sakai.
通讯作者：
Takafumi Sakai.

Localization of Ghrelin-Producing cells in the stomach of the rainbowtrout (Oncorhynchus mykiss).

虹鳟鱼 (Oncorhynchus mykiss) 胃中生长素释放肽生成细胞的定位。

DOI：
发表时间：
2004
期刊：
Zoological Science 21
影响因子：
0
作者：
Ichiro Sakata;Tsukasa Mori;Hiroyuki Kaiya;Mami Yamzki;Kenji Kanagwa;Kinji Inoue;Takafumi Sakai
通讯作者：
Takafumi Sakai