Transposon-mediated gene trapping in zebrafish
Transposon-mediated gene trapping in zebrafish
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DOI：
10.1016/j.ymeth.2005.12.006
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发表时间：
2006-07-01
期刊：
影响因子：
4.8
通讯作者：
Kawakami, Koichi
中科院分区：
文献类型：
--
作者：
Kotani, Tomoya;Nagayoshi, Saori;Kawakami, Koichi
Tol 2转座子系统可以非常有效地在斑马鱼生殖谱系中产生染色体插入。我们构建了一个基于Tol 2的基因捕获载体T2 KSAG，它包含一个剪接受体，GFP基因和polyA信号。在使用T2 KSAG进行基因捕获的中试筛选中，我们鉴定了38条在特定器官和组织中表达GFP的鱼品系。在SAGp 53 A系中，GFP在前脑和中脑中表达，并且基因捕获构建体的插入捕获了编码人KARP（Ku 86自身抗原相关蛋白）结合蛋白（KAB）的斑马鱼同源物的kab基因的转录物。在SAGm 18 B系中，GFP在中枢神经系统中表达，插入捕获了琥珀酰CoA：3-含氧酸CoA转移酶（SCOT）基因的转录本。在这里，我们描述了我们如何进行基因陷阱筛选和表征基因陷阱插入，并将讨论试点筛选的结果。(c)2006爱思唯尔公司All rights reserved.
The Tol2 transposon system can create chromosomal insertions in the zebrafish germ lineage very efficiently. We constructed a Tol2-based gene trap vector, T2KSAG, which contains a splice accepter, the GFP gene and the polyA signal. In the pilot screen for gene trapping using T2KSAG, we identified 38 fish lines expressing GFP in specific organs and tissues. In the SAGp53A line, GFP is expressed in the forebrain and midbrain, and the insertion of the gene trap construct captured a transcript of the kab gene encoding a zebrafish homolog of the human KARP (Ku86 autoantigen related protein)-binding protein (KAB). In the SAGm18B line, GFP is expressed in the central nervous system, and the insertion captured a transcript of a gene for succinyl CoA:3-oxoacid CoA-transferase (SCOT). Here, we describe how we performed the gene trap screen and characterized the gene trap insertions and will discuss the outcome of the pilot screen. (c) 2006 Elsevier Inc. All rights reserved.