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异齿裂腹鱼PYY基因的克隆、组织分布及其调节摄食功能研究
商振达, 刘锁珠, 谭占坤,等
西藏农牧学院 动物科学学院,西藏高原饲料加工工程研究中心
摘要:
【目的】克隆异齿裂腹鱼肽YY基因(PYY)的编码区(CDS)全长序列,对其生物信息学进行分析,并研究PYY基因在异齿裂腹鱼组织中的分布情况,以及餐前餐后和禁食复喂对PYY基因表达量的影响,为异齿裂腹鱼摄食调控机制研究提供依据。【方法】以采集于西藏雅鲁藏布江的7~12龄,体质量为(1.22±0.20) kg/尾的异齿裂腹鱼为对象,采用RT-PCR和Race-PCR技术克隆异齿裂腹鱼PYY基因的CDS全长序列,分析其编码蛋白质的氨基酸序列和蛋白结构;采用实时荧光定量PCR,分析PYY基因在异齿裂腹鱼组织中的分布情况,以及餐前餐后和禁食复喂对PYY基因在异齿裂腹鱼脑组织中表达量的影响。【结果】异齿裂腹鱼PYY基因CDS序列长度为735 bp,其中开放阅读框(ORF)为294 bp,编码98个氨基酸。系统进化分析表明,异齿裂腹鱼PYY基因与同属裂腹鱼属的鱼类亲缘关系最近;氨基酸序列分析表明,PYY蛋白质包括典型的特征序列和信号肽,其蛋白结构中具有亲水性结构和跨膜结构。PYY基因在异齿裂腹鱼脑组织中高度表达,在其他组织中微弱表达;餐前异齿裂腹鱼PYY基因的表达量无显著性差异,但餐后PYY基因的表达量显著升高,且随着餐后时间的逐渐延长,其表达量继续升高;禁食使异齿裂腹鱼脑组织中的PYY基因表达量下降,复喂后PYY基因的表达量极显著升高。【结论】成功克隆了异齿裂腹鱼PYY基因的CDS全长序列,PYY基因在异齿裂腹鱼脑组织中高度表达,但在肠道中微量表达,异齿裂腹鱼PYY基因是餐后饱感信号因子,具有调节动物摄食的功能。
关键词:  异齿裂腹鱼  肽YY  基因克隆  组织分布  摄食功能
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基金项目:西藏自治区厅校联合基金项目(XZ2017ZRG-12(Z));西藏高原饲料加工工程研究中心项目(XZJYT2018GCZX);中央财政支持地方高校发展专项资金项目(ZZXT2019-02)
Cloning,tissue distribution and regulating appetite function of PYY in Schizothorax o’connori
SHANG Zhenda, LIU Suozhu, TAN Zhankun,et al
Abstract:
【Objective】This study cloned Schizothorax o’connori PYY gene,analyzed its bioinformatics,and detected tissue distribution and mRNA levels of PYY gene before and after feeding,fasting,and refeeding to provide basis for understanding appetite regulatory system of CCK inSchizothorax o’connori.【Method】Schizothorax o’connori at the age of 7-12 years with weight of (1.22±0.20) kg/tail were collected from Yarlung Zangbo River in Tibet.The full-length CDS sequence of Schizothorax o’connori PYY gene was cloned by RT-PCR and Race-PCR technology,and amino acid sequence and protein structure were analyzed.The tissue distribution and appetite regulation of PYY in Schizothorax o’connori was investigated using real-time PCR technology.【Result】PYY gene sequence of Schizothorax o’connori was 735 bp and the open reading frame (ORF) sequence was 294 bp,encoding 98 amino acids.Phylogenetic analysis indicated that the PYY gene had the highest homology with Schizothorax.Amino acid sequence analysis showed that the PYY gene included typical characteristic sequence and signal peptide,and its protein structure had hydrophilic structure and transmembrane structure.According to the tissue distribution study,the PYY gene was highly expressed in brain and weakly expressed in other tissues.There was no significant difference in the expression of PYY gene before feeding,but the expression of PYY gene increased significantly after feeding,and its expression increased gradually with the extension of postprandial time.Fasting reduced the expression of PYY gene in brain and refeeding increased the expression of PYY gene.【Conclusion】This study successfully cloned the full length CDS sequence of Schizothorax o’connori PYY gene.It was highly expressed in brain and weakly expressed in intestine.The PYY gene was the signal factor for postprandial satiety with the function of regulating appetite.
Key words:  Schizothorax o’connori  peptide YY  molecular cloning  tissue distribution  appetite regulation