Heat-inducible expression of a reporter gene detected by transient assay in zebrafish
Heat-inducible expression of a reporter gene detected by transient assay in zebrafish
复制标题
DOI：
10.1006/excr.2000.4805
复制
发表时间：
2000-04-10
影响因子：
3.7
通讯作者：
Orbán, L
中科院分区：
文献类型：
--
作者：
Adám, A;Bártfai, R;Orbán, L
利用瞬时表达系统研究了两种表达LacZ基因的报告载体在小鼠和非洲爪哇HSP70启动子控制下在斑马鱼(Danio Rerio)胚胎中的热诱导活性。用组织化学方法将表达β-半乳糖苷酶的细胞染成蓝色，以每个胚胎的平均数量作为报告基因表达水平的指标。这两个构建体在胚胎组织中都是热诱导的，并表现出类似的热依赖性(表达水平从35-36摄氏度增加到39摄氏度，40摄氏度时明显下降)，类似于斑马鱼HSP70基因。然而，它们的诱导动力学不同，这可能是由于它们的5‘UTRs不同所致。两个HSP/LacZ构建体和一个内源HSP70基因的空间表达模式在RNA水平上基本相似。这些结果表明，我们的方法适用于体内热休克反应的分析，外源热休克启动子可能有助于诱导转基因在鱼类中的表达。(C)2000年学术出版社。
Heat-inducibility of two reporter constructs expressing lacZ gene under the control of mouse and Xenopus hsp70 promoters was tested in zebrafish (Danio rerio) embryos using a transient expression system. Cells expressing p-galactosidase were stained blue by histochemical staining and their average number per embryo was used as an indicator of the expression level of the reporter gene. Both constructs were heat-inducible in the embryonic tissues and showed similar heat dependence (increasing expression levels from 35-36 degrees C up to 39 degrees C with an apparent decrease at 40 degrees C), resembling that of the zebrafish hsp70 genes. However, their induction kinetics were different, which might be due to differences in their 5' UTRs. Spatial expression patterns of the two hsp/lacZ constructs and an endogenous hsp70 gene were mostly similar on the RNA level. These results indicate that our approach is applicable for in vivo analysis of the heat-shock response and that exogenous heat-shock promoters may be useful for inducible expression of transgenes in fish. (C) 2000 Academic Press.