Generation of Ci-Brachyuty-GFP stable transgenic lines in the ascidian Ciona savignyi
Generation of Ci-Brachyuty-GFP stable transgenic lines in the ascidian Ciona savignyi
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DOI：
10.1002/gene.10195
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发表时间：
2003-04-01
期刊：
影响因子：
1.5
通讯作者：
Smith, WC
中科院分区：
文献类型：
--
作者：
Deschet, K;Nakatani, Y;Smith, WC
我们报道了一批携带Ci-brachyury启动子/绿色荧光蛋白报告基因(Ci-Bra-GFP)的海鞘转基因品系的建立。将L-SCEL内切酶与含有L-SCEL侧翼识别位点的转基因载体共注射到受精卵中，获得转基因品系。在测试的12只F-0成虫中，有两只创始动物被发现将转基因传递给他们的后代(F(1)S)，频率分别为42%和23%。转基因进一步被F-2以孟德尔方式遗传，并表现出非镶嵌表达，表明整合到基因组中。孟德尔式的遗传和马赛克的缺失一直延续到F-3和F-4世代。Southern杂交分析表明，该转基因以不超过10个拷贝的串联阵列形式存在。利用这些Ci-Bra-GFP转基因基因，我们描述了野生型和突变型胚胎中涉及脊索形态发生的细胞运动和形状变化。(C)2003年Wiley-Liss，Inc.
We report generation of stable transgenic lines of the ascidian Ciona savignyi carrying a Ciona intestinalis-Brachyury-promoter/Green Fluorescent Protein-reporter (Ci-Bra-GFP) construct. The transgenic lines were made using a technique in which the endonuclease l-Scel was coinjected into fertilized eggs with a transgene construct containing flanking recognition sites for l-Scel. Two founder animals, out of 12 F-0 adults tested, were found to transmit the transgene to their offspring (F(1)s) at frequencies of 42% and 23%. The transgene was further inherited by the F-2 in a Mendelian fashion and displayed nonmosaic expression, indicating integration into the genome. The Mendelian inheritance and the absence of mosaicism persisted through the F-3 and F-4 generations. Southern blot analyses showed that the transgene was organized in tandem arrays of no more than 10 copies. Using these Ci-Bra-GFP transgenics, we describe cellular movements and shape changes involved in notochord morphogenesis in both wildtype and mutant embryos. (C) 2003 Wiley-Liss, Inc.