斑马鱼HCS基因全长cDNA序列及早期仔鱼阶段的空间表达

Cloning and expression of HCS gene in zebrafish larvae

蛋白生物素酰化发挥重要的生理作用,羧化酶和组蛋白生物素酰化主要依赖于羧化全酶合成酶(holocarboxylase synthetase,HCS)。首次克隆了HCS基因全长序列2 942 bp,其编码829个氨基酸,含有BirA功能结构域。利用RNA整体原位杂交技术,检测HCS基因在斑马鱼早期发育过程中的空间表达,发现HCS基因在脑、脊髓、鳃、心脏、肠道和肌肉等组织中表达。在肌肉组织中,HCS基因在初孵仔鱼的肌节表达,而在孵化后第6天仔鱼中,HCS基因则集中在每一肌节的中部表达。这些结果,为进一步了解HCS的功能奠定了基础。

Some proteins or enzymes play physiologic roles depending on its biotinylation via catalyzation of holocarboxylase synthetase （ HCS）, such as histones and carboxylases. So far, our understanding of HCS function is mainly from the researches on human cell lines. In order to understand the role of protein biotinylation in fish development, we cloned full-length HCS eDNA sequence with 2 942 bp. Intergrity open reading frame encodes 829 amino acids, containing conserved BirA ligase domain close to C-termino. Further investigation using whole mount RNA in situ hybridization shows that HCS gene is expressed in wide tissues such as brain, spine cord, gill, heart, intestine and sarcomere, indicating that it might play universal roles during organs development. It is interesting that HCS gene is expressed in whole sarcomere at new hatching larvae, whereas at 6 DAH （ day after hatching） larvae, expression signal of HCS gene concentrated on the middle of each sarcomere. Above results may be helpful for future studies on HCS function during fish development.