摘要
为探讨otx2基因在七彩神仙鱼(Symphysodon haraldi)亲代抚育过程中的作用,本试验采用RT-PCR法克隆得到了七彩神仙鱼otx2基因,otx2基因的cDNA序列的CDS区序列长度为873 bp,共编码290个氨基酸,预测分子质量为31771.55 Da,理论等电点为9.40。运用SMART、Swiss-model和MEGA软件对基因的结构及物种进化关系进行分析。氨基酸序列预测显示,该基因具有otx保守结构域,在进化关系上与尼罗罗非鱼和白边锯鳞鱼亲缘关系相近,同源性分析发现,其氨基酸序列与尼罗罗非鱼、长、西瓜刨、牙鲆和白边锯鳞鱼的同源性都在75%以上。荧光定量PCR分析显示,otx2基因在雌性七彩神仙鱼的脑、心脏、肝脏、肾脏等组织中都有表达,并且在脑组织中显著高表达。荧光原位杂交结果显示,otx2基因在七彩神仙鱼脑组织中的小球周灰质带、纵枕、顶盖室等区域均有表达。
This experiment used RT-PCR method to clone the otx2 gene of discus to explore the role of otx2 gene in the parental care process of discus(Symphysodon haraldi).The CDS region of otx2 cDNA sequence was 873 bp,290 amino acids were encoded,the predicted molecular mass was 31771.55 Da,and the theoretical isoelectric point was 9.40.SMART,Swiss-Model and MEGA software were used to analyze the gene structure and the relationship between species evolution.Amino acid sequence analysis showed that the gene had a conserved otx domain,which was close to Oreochromis niloticus and Myripristis murdjan in terms of evolutionary relationship.According to the homology analysis,the amino acid sequence of the gene was more than 75%homology with Oreochromis niloticus,Echeneis naucrates,Salarias fasciatus,Paralichthys olivaceus and Myripristis murdjan.Fluorescent quantitative PCR analysis showed that otx2 gene was expressed in the brain,heart,liver,kidney and other tissues of female discus,and the expression was significantly high in the brain tissues.In order to determine the location of otx2 gene in the brain tissue,fluorescence in situ hybridization was conducted in this experiment.The results of fluorescence in situ hybridization showed that otx2 gene was expressed in the peri-glomerular gray matter band,longitudinal occiput and parietal compartment in the brain tissue of the discus.This study provides primary data for exploring the role of otx2 gene in the parental care process of discus.
作者
魏域玲
温彬
高建忠
陈再忠
WEI Yu-ling;WEN Bin;GAO Jian-zhong;CHEN Zai-zhong(National Demonstration Center for Experimental Fisheries Science Education,Shanghai Ocean University,Shanghai 201306,China;Key Laboratory of Freshwater Aquatic Genetic Resources,Ministry of Agriculture and Rural Affairs,Shanghai Ocean University,Shanghai 201306,China;Shanghai Collaborative Innovation for Aquatic Animal Genetics and Breeding,Shanghai Ocean University,Shanghai 201306,China;Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306,China)
出处
《淡水渔业》
CSCD
北大核心
2021年第3期13-19,共7页
Freshwater Fisheries
基金
上海市自然科学基金(20ZR1423600)。
