Expression of a vas::EGFP transgene in primordial germ cells of the zebrafish
Expression of a vas::EGFP transgene in primordial germ cells of the zebrafish
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DOI：
10.1016/s0925-4773(02)00154-5
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发表时间：
2002-08-01
影响因子：
2.6
通讯作者：
Olsen, LC
中科院分区：
文献类型：
--
作者：
Krovel, AV;Olsen, LC
在斑马鱼中，母体产生的vasa（vas）转录物被靶向到早期胚胎的卵裂平面，随后被掺入原始生殖细胞（PGCs）。合子输精管转录发生在原肠胚形成开始时。在这里，我们报告的斑马鱼输精管轨迹的表征。该基因由27个外显子组成，跨度约25 kb，含有两个富含CpG的区域。我们利用vas调控区建立了转基因斑马鱼品系，在其原始生殖细胞中表达增强型绿色荧光蛋白（EGFP）。母体编码的vas：：EGFP转录物和VAS：：EGFP蛋白在胚胎发生过程中与PGCs分离。我们发现母体沉积的vas：：EGFP转录本在胚胎发生过程中至少在50 It的发育过程中是稳定的。Vas：：EGFP转录本不能在从雄性遗传转基因的胚胎中检测到，这很可能是由于缺乏早期合子表达所需的一个或多个调控元件。我们发现，vas：：EGFP转录本在早期胚胎的卵裂平面变得丰富，这一发现支持RNA定位信号定位在这些转录本的vas区域内。(C)2002爱思唯尔科学爱尔兰有限公司保留所有权利。
In zebrafish, maternally produced vasa (vas) transcripts become targeted to the cleavage planes of early embryos and subsequently incorporated into the primordial germ cells (PGCs). Zygotic vas transcription occurs from the onset of gastrulation. Here, we report on the characterisation of the zebrafish vas locus. The gene consists of 27 exons, spans about 25 kb, and contains two CpG-rich regions. We have used vas regulatory regions to establish transgenic zebrafish lines expressing enhanced green fluorescent protein (EGFP) in their PGCs. Maternally encoded vas::EGFP transcripts and VAS::EGFP protein segregate with the PGCs during embryogenesis. We find that the maternally deposited vas::EGFP transcripts are stable during embryogensis at least up to 50 It of development. Vas::EGFP transcripts could not be detected in embryos that inherit the transgene from males, most likely due to the lack of one or more regulatory elements required for early zygotic expression. We show that vas::EGFP transcripts become enriched to the cleavage planes in early embryos, a finding that supported an RNA localisation signal localised within the vas region of these transcripts. (C) 2002 Elsevier Science Ireland Ltd. All rights reserved.