3xP3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards
3xP3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards
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DOI：
10.1016/s0965-1748(01)00150-3
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发表时间：
2002-03-01
影响因子：
3.8
通讯作者：
Chavancy, G
中科院分区：
文献类型：
--
作者：
Thomas, JL;Da Rocha, M;Chavancy, G
最近在家蚕中实现了转基因，但是已经证明筛选大量后代以鉴定转基因个体是困难和耗时的。由于3xP 3-EGFP标记已被证明是转基因昆虫的合适的通用标记（Nature 402（1999）370），我们评估了其用于胚胎阶段筛选B的用途。Mori L.生殖系转化使用piggyBac衍生的载体pBac{3xP 3-EGFPaf}，我们能够从约120，000个胚胎（560窝）中分离出4个转基因个体。由于EGFP在G1胚胎干细胞中产生，通过半透明的卵绒毛膜可以看到，因此筛选是直接的。从胚胎发育的第五天起，EGFP就在干细胞以及中枢和外周神经系统中产生。在整个幼虫期，它在茎中以高水平持续存在，并且还存在于蛹的复眼和神经组织以及蛾的复眼中。(C)2002爱思唯尔科技有限公司版权所有。
Transgenesis was recently achieved in Bombyx mori L., but it has proved difficult and time-consuming to screen the numerous progeny to identify the transgenic individuals. As the 3xP3-EGFP marker has been shown to be a suitable universal marker for transgenic insects (Nature 402 (1999) 370), we evaluated its use for embryonic-stage screening for B. mori L. germline transformation. Using the piggyBac-derived vector pBac{3xP3-EGFPaf}, we were able to isolate four transgenic individuals from about 120,000 embryos (560 broods). The screening was straightforward due to EGFP production in the G1 embryonic stemmata, which was visible through the translucent egg chorion. EGFP was produced in the stemmata and central and peripheral nervous systems from the fifth day of embryonic development. It persisted at high levels in the stemmata throughout the larval stage, and was also present in the compound eyes and nervous tissues of the pupae and the compound eyes of the moths. (C) 2002 Elsevier Science Ltd. All rights reserved.