O'Boyle Shaun, Bree Ronan T, McLoughlin Sarah, Grealy Maura, Byrnes Lucy
Department of Biochemistry and National Centre for Biomedical Engineering Science, National University of Ireland, Galway, Ireland.
Biochem Biophys Res Commun. 2007 Jun 29;358(2):462-8. doi: 10.1016/j.bbrc.2007.04.116. Epub 2007 Apr 26.
Early development of the embryo is directed by maternal gene products and characterised by limited zygotic gene activity, cell division synchrony and no cell motility in several vertebrates including fish and frogs. At the midblastula transition (MBT), zygotic transcription is grossly activated, cells become motile and cell divisions become asynchronous. The aim of this study was to identify genes whose expression is up-regulated at the MBT in zebrafish. Suppression subtractive hybridisation (SSH) was employed to isolate 48 unique cDNAs, 28 of which show significant similarity to known genes and 20 represent novel cDNAs. Twenty one of these genes, with potential roles in transcriptional regulation, cell cycle control, and embryonic patterning showed increased expression at the MBT. Our results demonstrate the value of SSH as a tool to clone novel, zygotic, developmentally regulated genes that may be important in the progression of the MBT and embryonic patterning.
胚胎的早期发育由母体基因产物主导,其特征是包括鱼类和青蛙在内的几种脊椎动物的合子基因活性有限、细胞分裂同步且无细胞运动。在中囊胚转换(MBT)时,合子转录被大量激活,细胞开始运动且细胞分裂变得不同步。本研究的目的是鉴定在斑马鱼MBT时表达上调的基因。采用抑制性消减杂交(SSH)技术分离出48个独特的cDNA,其中28个与已知基因有显著相似性,20个代表新的cDNA。这些基因中有21个在转录调控、细胞周期控制和胚胎模式形成中具有潜在作用,它们在MBT时表达增加。我们的结果证明了SSH作为一种工具在克隆新的、合子的、受发育调控的基因方面的价值,这些基因可能在MBT进程和胚胎模式形成中起重要作用。