斑马鱼丝氨酸羟甲基转移酶1基因的生物信息学和表达分析  

Bioinformatics and Expression Analysis of Serine Hydroxymethyltransferase 1 Gene in Zebrafish

作　　者：陈炳宇 何兴 夏佳敏 邹蓉 黄四洲[1,2] Chen Bingyu;He Xing;Xia Jiamin;Zou Rong;Huang Sizhou(Department of Human Anatomy,Histology and Embryology,Chengdu Medical College,Chengdu 610500,China;Key Laboratory of Development and Regeneration of Sichuan Province,Chengdu 610500,China;School of Biomedical Sciences,Chengdu Medical College,Chengdu 610500,China)

机构地区：[1]成都医学院人体解剖与组织胚胎学教研室,成都610500 [2]发育与再生四川省重点实验室,成都610500 [3]成都医学院生物医学院,成都610500

出　　处：《成都医学院学报》2021年第6期681-687,共7页Journal of Chengdu Medical College

基　　金：国家自然科学基金(No:32070805);四川省科技厅基础研究项目(No:20YYJC3369);成都医学院创新团队项目(No:CYTD19-01);发育与再生四川省重点实验室项目(No:20180498)。

摘　　要：目的探索斑马鱼丝氨酸羟甲基转移酶1(shmt1)基因的生物学结构和基因表达模式。方法利用生物信息学方法对斑马鱼shmt1的基因结构、物种间同源性、物种间进化、蛋白质二级结构和三级结构等进行分析和预测,并用整胚原位杂交技术探究斑马鱼shmt1在重要器官及胚胎早期发育中的表达。结果斑马鱼shmt1位于3号染色体(ZDB-GENE-040426-1558)上,全长15 142 bp,能编码481个氨基酸,属于天冬氨酸转氨酶(AAT)超家族,其二级结构以α-螺旋和无规则卷曲为主,且斑马鱼shmt1蛋白与其他哺乳动物同源物在进化上高度保守。反转录定量PCR(RT-PCR)结果显示,斑马鱼shmt1不仅在胚胎发育的各个时期表达,而且也在成年鱼的肾脏、心脏和性腺中高表达。原位杂交结果显示,shmt1除了在卵黄多核层、鼻翼板中脑区、视顶盖和视泡等处集中表达,还在肝脏特异性表达。结论 shmt1作为一碳代谢叶酸循环中的重要蛋白,在斑马鱼发育早期的胚胎卵黄多核层、鼻翼板中脑区、视顶盖和视泡、肝脏特异性表达。该研究为今后以斑马鱼为模型动物探究shmt1基因在器官发育中的功能研究奠定基础。Objective To study the biological structure and gene expression pattern of serine hydroxymethyltransferase 1(shmt1) gene in zebrafish.Methods The gene structure, interspecies homology, interspecies evolution, protein secondary structure and protein tertiary structure of zebrafish shmt1 were predicted and analyzed by bioinformatics methods. And the expression of shmt1 in vital organs and early embryonic development of zebrafish was investigated by whole embryo in situ hybridization. Results Zebrafish shmt1 is located on chromosome 3(ZDB-GENE-040426-1558), with a total length of 15142 bp, encoding 481 amino acids, belonging to the aspartate aminotransferase(AAT) superfamily, with its secondary structure dominated by alpha-helix and irregular crimp. And the zebrafish shmt1 protein and other mammalian homolog are highly conserved in evolution. Reverse transcription-polymerase chain reaction(RT-PCR) results showed that zebrafish shmt1 was not only expressed in various stages of embryonic development, but also highly expressed in kidney, heart and gonad of adult fish. The results of in situ hybridization showed that shmt1 was expressed specifically in the liver in addition to the concentrated expression of the yolk polynuclear layer, the midbrain area of the nasal alar plate, the optic tectum, and the optic vesicles.Conclusion As an important protein in carbon metabolism and folate cycle, shmt1 is specifically expressed in the yolk polynuclear layer, the midbrain area of the nasal alar plate, the optic tectum, and the optic vesicles in the early stage of zebrafish development. This study will lay a foundation for future studies on the function of shmt1 gene in organ development with zebrafish as the model animal.

关 键 词：丝氨酸羟甲基转移酶1 斑马鱼 肝脏 代谢 

分 类 号：Q71[生物学—分子生物学]