Transient and stable GFP expression in germ cells by the vasa regulatory sequences from the red seabream (Pagrus major).
Transient and stable GFP expression in germ cells by the vasa regulatory sequences from the red seabream (Pagrus major).
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红鲷（真鲷）的 vasa 调控序列在生殖细胞中瞬时和稳定 GFP 表达
DOI：
10.7150/ijbs.4421
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发表时间：
2012
影响因子：
9.2
通讯作者：
Hong Y
中科院分区：
文献类型：
--
作者：
Lin F;Liu Q;Li M;Li Z;Hong N;Li J;Hong Y
原始生殖细胞（PGCs）是负责遗传传递给下一代的配子的前体。它们为生物多样性的低温保存和恢复提供了理想的系统。近年来，对PGCs在种内和种间的可视化、分离和移植研究引起了人们的广泛关注。在鱼类中，在活胚胎和个体中稳定的PGC可视化仅限于实验鱼模型，如medaka和斑马鱼。一个例外是虹鳟鱼，它代表了唯一具有水产养殖重要性的物种，并且在整个发育过程中都有gfp标记的生殖细胞。通过胚胎注射含有绿色荧光蛋白基因（GFP）和母胚基因vasa、nos1等3′-非翻译区（3′-UTR）的mRNA，可以对PGCs进行瞬时标记。利用胚基因的一些转录调控序列，如vasa启动子和3′-UTR，可以通过生产转基因动物来实现稳定的PGC标记。在这项研究中，我们报道了红海鲷vasa （Pmvas）调控序列的功能分析，以medaka为模型系统。结果表明，注射GFP-Pmvas3'UTR mRNA能够在胚胎发生过程中标记medaka PGCs。此外，我们构建了pPmvasGFP转基因载体，并在Pmvas转录调控序列的控制下，建立了在生殖细胞（包括PGCs、有丝分裂生殖细胞和减数分裂生殖细胞）中稳定表达GFP的转基因medaka细胞系。结论：本研究检测的Pmvas调控序列足以用于生殖细胞的表达和标记。
Primordial germ cells (PGCs) are the precursors of gametes responsible for genetic transmission to the next generation. They provide an ideal system for cryopreservation and restoration of biodiversity. Recently, considerable attention has been raised to visualize, isolate and transplant PGCs within and between species. In fish, stable PGC visualization in live embryo and individual has been limited to laboratory fish models such as medaka and zebrafish. One exception is the rainbow trout, which represents the only species with aquaculture importance and has GFP-labeled germ cells throughout development. PGCs can be transiently labeled by embryonic injection of mRNA containing green fluorescence protein gene (GFP) and 3'-untranslated region (3'-UTR) of a maternal germ gene such as vasa, nos1, etc. Stable PGC labeling can be achieved through production of transgenic animals by some transcriptional regulatory sequences from germ genes, such as the vasa promoter and 3'-UTR. In this study, we reported the functional analyses of the red seabream vasa (Pmvas) regulatory sequences, using medaka as a model system. It was showed that injection of GFP-Pmvas3'UTR mRNA was able to label medaka PGCs during embryogenesis. Besides, we have constructed pPmvasGFP transgenic vector, and established a stable transgenic medaka line exhibiting GFP expression in germ cells including PGCs, mitotic and meiotic germ cells of both sexes, under control of the Pmvas transcriptional regulatory sequences. It is concluded that the Pmvas regulatory sequences examined in this study are sufficient for germ cell expression and labeling.
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影响因子：
1.2
作者：
Liu, Lixiu;Hong, Ni;Hong, Yunhan
通讯作者：
Hong, Yunhan
影响因子：
0.7
作者：
Goto-Kazeto, Rie;Saito, Taiju;Yamaha, Etsuro
通讯作者：
Yamaha, Etsuro
影响因子：
0.7
作者：
Saito, Taiju;Fujimoto, Takafumi;Yamaha, Etsuro
通讯作者：
Yamaha, Etsuro
影响因子：
2.5
作者：
Kobayashi, T;Yoshizaki, G;Takeuchi, T
通讯作者：
Takeuchi, T
影响因子：
4
作者：
Fan, Lianchun;Moon, Jesung;Collodi, Paul
通讯作者：
Collodi, Paul