Department of Biological Sciences, National University of Singapore, Singapore, Singapore 117543.
Mar Biotechnol (NY). 2010 Feb;12(1):1-7. doi: 10.1007/s10126-009-9217-y. Epub 2009 Jul 10.
A stable Tg(UAS:GFP) zebrafish line was generated and crossed with Tg(hsp70:GAL4) line, in which the GAL4 gene is under the control of an inducible zebrafish promoter derived from the heat shock 70 protein gene (hsp70). The dynamic green fluorescent protein (GFP) expression in early zebrafish embryos in the GAL4/UAS binary system was then investigated. We found that, at early developmental stages, expression of GFP effector gene was restricted and required a long recovery time to reach a detectable level. At later developmental stage (after 2 days postfertilization), GFP could be activated in multiple tissues in a shorter time, apparently due to a higher level of GAL4 messenger RNA induction. It appears that the type of tissues expressing GFP was dependent on whether they had been developed at the time of heat shock. Therefore, the delayed and restricted transgene expression should be taken into consideration when GAL4/UAS system is used to study transgene expression in early developmental stages.
我们构建了一个稳定的 Tg(UAS:GFP) 斑马鱼品系，并与 Tg(hsp70:GAL4) 线杂交，其中 GAL4 基因受来自热休克 70 蛋白基因（hsp70）的诱导型斑马鱼启动子的控制。然后研究了 GAL4/UAS 双元系统中早期斑马鱼胚胎中绿色荧光蛋白（GFP）的动态表达。我们发现，在早期发育阶段，GFP 效应基因的表达受到限制，需要较长的恢复时间才能达到可检测水平。在较晚的发育阶段（受精后 2 天），GFP 可以在较短的时间内在多个组织中被激活，这显然是由于 GAL4 mRNA 的诱导水平更高。似乎表达 GFP 的组织类型取决于它们在热休克时是否已经发育。因此，当使用 GAL4/UAS 系统研究早期发育阶段的转基因表达时，应考虑到转基因表达的延迟和限制。
