鳙goat基因序列特征及其时空表达模式分析  

Analysis of sequence characterization and spatiotemporal expression pattern of goat gene in bighead carp(Hypophthalmichthys nobilis)

作　　者：王盼[1] 傅建军[2] 束世长 朱文彬[2] 罗明坤 王兰梅[2] 董在杰 WANG Pan;FU Jian-jun;SHU Shi-chang;ZHU Wen-bin;LUO Ming-kun;WANG Lan-mei;DONG Zai-jie(Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources Ministry of Education(Shanghai Ocean University),Shanghai 201306,China;Freshwater Fisheries Research Center,Chinese Academy of Fishery Sciences/Key Laboratory of Freshwater Fisheries and Germplasm Resource Utilization,Ministry of Agriculture and Rural Affairs,Wuxi,Jiangsu 214081,China)

机构地区：[1]水产种质资源发掘与利用教育部重点实验室(上海海洋大学),上海201306 [2]中国水产科学研究院淡水渔业研究中心/农业农村部淡水渔业和种质资源利用重点实验室,江苏无锡214081

出　　处：《南方农业学报》2023年第9期2533-2543,共11页Journal of Southern Agriculture

基　　金：国家现代农业产业技术体系建设专项(CARS-45-05)。

摘　　要：【目的】探究鳙的ghrelin O-acyltransferase(goat)基因序列特征及时空表达模式,解析goat基因的潜在生物学功能。【方法】通过RACE扩增得到goat基因cDNA全长序列,采用实时荧光定量PCR对其在早期发育阶段、各组织和冬夏季4个组织(脑、肠道、肌肉和肝脏)中的表达特征进行检测;结合原位杂交和免疫组化技术分别从基因和蛋白水平解析goat基因在肠组织中的分布特征,并通过实时荧光定量PCR评估goat基因及通路相关基因的共表达模式。【结果】鉴定得到goat基因cDNA序列全长2255 bp,开放阅读框(ORF)长1116 bp,共编码371个氨基酸残基。推导氨基酸序列含7个跨膜结构域,并包含参与催化的天冬酰胺(第251位)和组氨酸(第282位)。同源比对分析发现鳙GOAT氨基酸序列与鲤科鱼类相似度极高。鳙goat基因在早期孵化时和出膜后5 d的表达水平均显著高于其他时期(P<0.05,下同);成体组织差异表达分析发现goat基因在肠道中表达量最高;goat基因在夏季脑、肠道和肌肉组织的相对表达量均显著高于冬季。原位杂交试验结果显示goat基因在肠道中表达信号较强,且肠道切片的免疫组化试验显示肠黏膜中存在GOAT与GHRL共定位的阳性信号。goat基因与生长激素的合成、分泌和作用通路(ko04935)中相关基因存在共表达关系。【结论】goat基因具有MBOAT家族的典型结构特征,可能通过与摄食相关基因(ghrl等)协作来参与鳙摄食调控和生长代谢适应等生物学过程。【Objective】The purpose of the study was to investigate the sequence characterization and spatiotemporal expression pattern of ghrelin O-acyltransferase(goat)gene of bighead carp(Hypophthalmichthys nobilis),and to analyze the potential biological functions of goat gene.【Method】The full-length sequence of goat gene cDNA was obtained using the RACE amplification,and its expression characteristics in early developmental stages,different tissues and four tissues(brain,intestine,muscle and liver)in summer and winter were detected by real-time fluorescence quantitative PCR(qRT-PCR).The distribution of the goat gene in the intestine tissue was characterized through in situ hybridization and immuno-cytochemistry at the gene and protein levels,respectively.Moreover,co-expression patterns of goat gene and pathway-related genes were assessed by qRT-PCR.【Result】The cDNA sequence of the goat gene was identified as 2255 bp in length,with an open reading frame(ORF)of 1116 bp,encoding 371 amino acids residues.The deduced amino acid se-quence contained seven transmembrane regions and contained asparagine(the 251st position)and histidine(the 282nd posi-tion),which were involved in catalysis.Homology analysis revealed that goat gene of bighead carp was very similar to that of Cypriniformes species.The goat gene expression levels at the time of early hatching stage and 5 d after membrane emergence were significantly higher than those at other periods(P<0.05,the same below).Differential expression analy-sis of adult tissues found that goat gene expression was the highest in the intestine.The goat gene expression in brain,in-testine and muscle tissues was significantly higher in summer than in winter.Furthermore,in situ hybridization experi-ments revealed that goat gene was expressed with a strong signal in the intestine,and immunohistochemistry in intestinal sections showed the presence of positive signals of GOAT co-localized with GHRL in the intestinal mucosa.There was a co-expression relationship between goat gene and r

关 键 词：鳙 ghrelin O-acyltransferase基因(goat) 时空表达 摄食调控 生长代谢 

分 类 号：S965.114[农业科学—水产养殖]