zTrap: zebrafish gene trap and enhancer trap database.
zTrap: zebrafish gene trap and enhancer trap database.
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DOI：
10.1186/1471-213x-10-105
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发表时间：
2010-10-18
影响因子：
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通讯作者：
Yoshida M
中科院分区：
文献类型：
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作者：
Kawakami K;Abe G;Asada T;Asakawa K;Fukuda R;Ito A;Lal P;Mouri N;Muto A;Suster ML;Takakubo H;Urasaki A;Wada H;Yoshida M
我们已经开发了遗传方法在斑马鱼中使用Tol 2转座因子，即转基因，基因捕获，增强子捕获和Gal 4FF-UAS系统。基因陷阱构建体含有剪接受体和GFP或Gal 4FF（酵母Gal 4转录激活因子的修饰形式）基因，增强子陷阱构建体含有斑马鱼hsp 70 1启动子和GFP或Gal 4FF基因。通过使用这些构建体进行遗传筛选，我们已经产生了在特定细胞、组织和器官中表达GFP和Gal 4FF的转基因斑马鱼。通过产生携带Gal 4FF转基因和置于Gal 4识别序列（UAS）下游的GFP报告基因的双转基因鱼来可视化Gal 4FF表达。此外，可以通过与UAS效应鱼交配来操纵Gal 4FF表达细胞。例如，当在特定神经元中表达Gal 4FF的鱼与携带UAS下游的破伤风神经毒素基因的UAS：TeTxLC鱼杂交时，双转基因鱼中的神经元活动被抑制。因此，这些转基因鱼可用于发育生物学和神经生物学的研究。为了提高转基因鱼类资源的利用率，我们开发了一个基于Web的数据库zTrap http://kawakami.lab.nig.ac.jp/ztrap/。zTrap数据库包含GFP和Gal 4FF表达模式的图像，以及基因陷阱和增强子陷阱构建体的整合位点周围的基因组DNA序列。整合位点通过内部Blat分析映射到Ensembl斑马鱼基因组上，并且可以在zTrap和Ensembl基因组浏览器上查看。此外，zTrap还配备了搜索这些数据以获得表达模式和感兴趣的基因组位点的功能。zTrap包含关于转基因鱼的信息，包括UAS报告和效应鱼。zTrap是一种有用的资源，用于发现以所需模式表达GFP和Gal 4FF的基因陷阱和增强子陷阱鱼品系，并发现位于感兴趣的基因内或附近的基因陷阱和增强子陷阱构建体的插入。这些转基因鱼可以用来观察胚胎发生过程中的特定细胞类型，操纵它们的功能，并发现新的基因和顺式调控元件。因此，zTrap应有助于利用转基因斑马鱼资源进行基因组学、发育生物学和神经生物学研究。
We have developed genetic methods in zebrafish by using the Tol2 transposable element; namely, transgenesis, gene trapping, enhancer trapping and the Gal4FF-UAS system. Gene trap constructs contain a splice acceptor and the GFP or Gal4FF (a modified version of the yeast Gal4 transcription activator) gene, and enhancer trap constructs contain the zebrafish hsp70l promoter and the GFP or Gal4FF gene. By performing genetic screens using these constructs, we have generated transgenic zebrafish that express GFP and Gal4FF in specific cells, tissues and organs. Gal4FF expression is visualized by creating double transgenic fish carrying a Gal4FF transgene and the GFP reporter gene placed downstream of the Gal4-recognition sequence (UAS). Further, the Gal4FF-expressing cells can be manipulated by mating with UAS effector fish. For instance, when fish expressing Gal4FF in specific neurons are crossed with the UAS:TeTxLC fish carrying the tetanus neurotoxin gene downstream of UAS, the neuronal activities are inhibited in the double transgenic fish. Thus, these transgenic fish are useful to study developmental biology and neurobiology. To increase the usefulness of the transgenic fish resource, we developed a web-based database named zTrap http://kawakami.lab.nig.ac.jp/ztrap/. The zTrap database contains images of GFP and Gal4FF expression patterns, and genomic DNA sequences surrounding the integration sites of the gene trap and enhancer trap constructs. The integration sites are mapped onto the Ensembl zebrafish genome by in-house Blat analysis and can be viewed on the zTrap and Ensembl genome browsers. Furthermore, zTrap is equipped with the functionality to search these data for expression patterns and genomic loci of interest. zTrap contains the information about transgenic fish including UAS reporter and effector fish. zTrap is a useful resource to find gene trap and enhancer trap fish lines that express GFP and Gal4FF in desired patterns, and to find insertions of the gene trap and enhancer trap constructs that are located within or near genes of interest. These transgenic fish can be utilized to observe specific cell types during embryogenesis, to manipulate their functions, and to discover novel genes and cis-regulatory elements. Therefore, zTrap should facilitate studies on genomics, developmental biology and neurobiology utilizing the transgenic zebrafish resource.