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Studies on vertebrate developmental genes by transposon-mediated gene trapping

转座子介导的基因捕获研究脊椎动物发育基因

基本信息

批准号：
15201041
负责人：
KAWAKAMI Koichi
金额：
$ 22.8万
依托单位：
National Institute of Genetics.
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2003
资助国家：
日本
起止时间：
2003 至 2005
项目状态：
已结题

项目摘要

(1)We injected a transposon-donor plasmid containing a To12 construct and mRNA encoding the transposase into zebrafish fertilized eggs and demonstrated that the transposon construct can be integrated in the genome of the germ cells very efficiently. This method is now widely used as a standard method to create transgenic fish.(2)We constructed the To12 construct containing a splice acceptor and the GFP reporter gene and created random integration of this construct in the zebrafish genome. When the splice acceptor trapped a transcript of an endogenous gene, the GFP gene is expressed in a pattern similar to that of the endogenous gene. This was the first successful demonstration of gene trapping in zebrafish.(3)We have been making efforts to distribute this useful methodology by publication of manuscripts, talks in international meetings, writing review articles, a training course and collaboration.(4)In the course of the transposon studies in zebrafish, we also found that the To12 transposon system is also active in mouse and Xenopus. To12 will be a useful gene transfer vector in these model vertebrates.

(1)We将含有To12构建体和编码转座酶的mRNA的转座子供体质粒注射到斑马鱼受精卵中，并证明转座子构建体可以非常有效地整合到生殖细胞的基因组中。这种方法现在被广泛用作创建转基因鱼的标准方法。(2)We构建了含有剪接受体和GFP报告基因的To12构建体，并在斑马鱼基因组中创建了该构建体的随机整合。当剪接受体捕获内源基因的转录物时，GFP基因以与内源基因相似的模式表达。这是第一次在斑马鱼中成功地展示基因捕获。(3)We一直在努力通过出版手稿、在国际会议上发表演讲、撰写评论文章、举办培训班和开展合作等方式传播这一有用的方法。(4)In在斑马鱼转座子研究的过程中，我们还发现To12转座子系统在小鼠和非洲爪蟾中也是活跃的。To12将是这些模式脊椎动物中有用的基因转移载体。

项目成果

期刊论文数量（57）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Fugu ESTs:: New resources for transcription analysis and genome annotation

DOI：
10.1101/gr.1691503
发表时间：
2003-12-01
期刊：
GENOME RESEARCH
影响因子：
7
作者：
Clark, MS;Edwards, YJK;Johnson, SL
通讯作者：
Johnson, SL

ゼブラフィッシュの時空間特異的な遺伝子発現

斑马鱼的时空特异性基因表达

DOI：
发表时间：
2004
期刊：
蛋白質核酸酵素 49
影响因子：
0
作者：
Satoshi Nakamura;Kazumasa Yamamoto;Kazuya Takeda;Shingo Kuroiwa;Norihide Kitaoka;Takeshi Yamada;Mitsunori Mizumachi;Takanobu Nishiura;Masakiyo Fujimoto;Akira Saso;Toshiki Endo;小谷友也
通讯作者：
小谷友也

The status, quality, and expansion of the NIH full-length cDNA project: The Mammalian Gene Collection (MGC)