Induction of Japanese flounder TNF promoter activity by lipopolysaccharide in zebrafish embryo
Induction of Japanese flounder TNF promoter activity by lipopolysaccharide in zebrafish embryo
复制标题
DOI：
10.1007/s10126-004-0403-7
复制
发表时间：
2005-05-01
影响因子：
3
通讯作者：
Aoki, T
中科院分区：
文献类型：
--
作者：
Yazawa, R;Hirono, I;Aoki, T
用显微注射法将含有牙鲆肿瘤坏死因子(TNF)2381-bp启动子区域和绿色荧光蛋白(GFP)启动子的重组载体导入斑马鱼受精卵。在非应激条件下，2个转基因斑马鱼品系在心脏和咽部周围表达了绿色荧光蛋白。当胚胎受到脂多糖(LPS)刺激时，GFP在整个斑马鱼胚胎表面都有表达，Western印迹分析还表明，当LPS浓度为140 mU g/ml时，GFP蛋白在斑马鱼胚胎中的表达水平高于0和70 mU g/ml时，刀豆蛋白A或佛波酯刺激斑马鱼胚胎时，GFP蛋白表达较弱，但两者诱导的GFP蛋白表达较强。GFP的表达在刺激后1h达到高峰，然后逐渐下降。这些结果表明，斑马鱼胚胎中受肿瘤坏死因子启动子调控的转录可能受内毒素识别系统的控制。
A recombinant plasmid containing the 2381-bp promoter region of Japanese flounder tumor necrosis factor (TNF) and green fluorescence protein (GFP) was introduced into zebrafish fertilized eggs by microinjection. GFP was expressed in 2 transgenic zebrafish lines in the heart and around the pharynx under unstressed condition. When embryos were exposed to lipopolysaccharide (LPS), GFP was expressed in the whole zebrafish embryonic surface, and Western blot analysis also showed that the level of the expressed GFP protein in zebrafish embryo was higher at an LPS concentration of 140 mu g/ml than at LPS concentrations of 0 and 70 mu g/ml. Stimulation with either concanavalin A or phorbol myristate acetate induced weak GFP expression, but stimulation with both of them induced strong expressiori similar to that induced by LPS. GFP expression peaked 1 hour after stimulation, then gradually decreased. These results indicate that transcription regulated by the Japanese flounder TNF promoter could be under the control of the LPS-recognition system in zebrafish embryos.