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基于聚合酶链式反应的无内含子斑马鱼无尾基因克隆

PCR-based cloning of an intronless zebrafish no tail gene.

作者信息

Yamakoshi Kimi, Shimoda Nobuyoshi

机构信息

PRESTO, Japan Science and Technology Corporation (JST), Institute for Genome Research, University of Tokushima, 3-18-15 Kuramoto-cho, Tokushima 770-8503, Japan.

出版信息

Biochem Biophys Res Commun. 2003 Jun 27;306(2):598-602. doi: 10.1016/s0006-291x(03)01007-6.

Abstract

The zebrafish no tail gene (ntl) is indispensable for the formation of the notochord and tail structure. Here, we show the presence of an intronless ntl gene in zebrafish, which we designated cryptail (ctl). ctl could not be found in any zebrafish genomic resources examined and was only just cloned by a PCR-based approach that relied on its lack of introns and homology to ntl. The amplifiable region of ctl was confined to the transcribed region of ntl. ctl thus appeared to have been generated by reverse transcription of ntl mRNA, like a processed pseudogene. ctl was very polymorphic even in individual fish, but had no missense mutations. This may suggest that ctl has a physiological function in zebrafish.

摘要

斑马鱼无尾基因(ntl)对于脊索和尾部结构的形成不可或缺。在此,我们展示了斑马鱼中存在一个无内含子的ntl基因,我们将其命名为隐尾基因(ctl)。在所检测的任何斑马鱼基因组资源中均未发现ctl,它是通过基于PCR的方法克隆得到的,该方法依赖于其缺乏内含子以及与ntl的同源性。ctl的可扩增区域局限于ntl的转录区域。因此,ctl似乎是由ntl mRNA的逆转录产生的,类似于一个加工过的假基因。即使在个体鱼类中,ctl也具有高度多态性,但没有错义突变。这可能表明ctl在斑马鱼中具有生理功能。

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