摘要
通过同源克隆和染色体步移对光倒刺鲃(Spinibarbus hollandi)黑素皮质素受体-5 (melanocortin receptor-5,mc5r)基因进行了克隆,应用生物信息学方法对mc5r 基因序列和推测的氨基酸序列进行分析,采用荧光定量PCR 技术研究其mRNA 的组织分布、日周期变化以及饥饿再投喂的表达情况。结果表明mc5r 基因长度为1 947 bp,含有1 个987 bp 的外显子,编码329 个氨基酸。通过序列同源比对及系统发育进化分析,光倒刺鲃mc5r 的氨基酸序列与鲤(Cyprinus carpio)、犀角金线鲅(Sinocyclocheilus rhinocerous)的mc5r 同源性达到98%。光倒刺鲃mc5r 在大脑中显著表达,在间脑、心脏的表达量次之,在中脑、小脑表达量较少,在肾脏、胃、肌肉、脾、肠、性腺、肝脏、延脑、眼中微量表达。光倒刺鲃mc5r mRNA 的日周期表达量随昼夜节律而变化,饥饿再投喂实验发现饥饿再投喂组鱼其脑中表达量会显著升高,随后会有下降趋势,表明mc5r 可能与光倒刺鲃的摄食调控有关。
In this study, melanocortin receptor-5 (mc5r) gene of Spinibarbus hollandi was cloned by homology cloning and genome walking method. Bioinformatics methods were used to analyze the gene sequence and deduced amino acid sequence of mc5r. The tissue distribution, diurnal change and starvation response of mRNA expression were detected by real-time PCR. The full length of mc5r gene was 1 947 bp with one exon of 987 bp encoding 329 amino acids. According to multiple sequences alignment and phylogeny evolution analysis, the homology of S. hollandi mc5r amino acid sequence with Cyprinus carpio and Sinocyclocheilus rhinocerous reached 98%. S. hollandi mc5r had significant expression in brain, followed by diencephalon and heart, relatively less in mesencephalon and cerebellum, and weak in kidneys, stomach, muscles, spleen, intestines, gonadal, liver, medulla oblongata and eye. The diurnal periodicity expression of mc5r mRNA changed with day and night regularly. In the starvation refeeding experiment, the expression of refeeding fish increased significantly after starvation, but then decreased. It is indicated that mc5r may have some effects on food intake regulation.
作者
周惠强
陈凯棱
舒琥
杨扬
钟东明
张明清
ZHOU Huiqiang;CHEN Kailing;SHU Hu;YANG Yang;ZHONG Dongming;ZHANG Mingqing(School of Life Science,Guangzhuo University,Guangzhou 510006,China)
出处
《南方水产科学》
CAS
CSCD
北大核心
2019年第4期99-106,共8页
South China Fisheries Science
基金
公益性行业(农业)重大科研专项(201303048号)
广东省科技计划项目(2014A020208145)
关键词
光倒刺鲃
黑素皮质素受体-5
克隆
日周期表达
饥饿响应
Spinibarbus hollandi
melanocortin receptor-5
clone
diurnal expression
starvation response
