| 摘要: |
| 【目的】克隆黄颡鱼神经肽Y基因(NPY),研究其在不同组织及在饥饿情况下脑组织中的表达情况，探讨其在黄颡鱼摄食活动中的作用。【方法】利用RT-PCR和RACE技术，克隆黄颡鱼NPY基因的cDNA序列全长，对其编码氨基酸进行生物信息学分析；利用半定量PCR和实时荧光定量PCR技术，对NPY基因在黄颡鱼成体不同组织(脑、心脏、肝脏、肾脏、脾脏、胃、肌肉、鳃、性腺)中的表达情况进行研究，并检测饥饿不同时间(0,24，48，72，96，120，144和168 h)以及饥饿168 h重新投饵1，3和5 h后黄颡鱼脑组织中 NPY表达水平的变化。【结果】黄颡鱼NPY基因cDNA序列全长772 bp，开放阅读框282 bp，编码93个氨基酸，其与瓦氏黄颡鱼同源性最高(97%)，与斑点叉尾鮰、建鲤、胭脂鱼、中华倒刺鲃的同源性分别为87%，72%，72%和70%。NPY基因在黄颡鱼成体组织脑、脾脏、肝脏、鳃、性腺中都有表达，而在心脏、肌肉、胃、肾脏中不表达，在脑中的表达量极显著高于其他组织(P ＜0.01)；在饥饿处理24~144 h时,随饥饿时间的增加,黄颡鱼脑组织中NPY表达量升高，但在饥饿168 h重新投饵3 h后,NPY表达量即可下降到正常水平。【结论】NPY基因在黄颡鱼脑中大量表达，随着饥饿时间的增加脑组织中NPYmRNA 表达量上升，重新投饵后其表达量很快下降到正常水平，表明NPY基因在黄颡鱼摄食活动中发挥着重要作用。 |
| 关键词: 黄颡鱼 NPY基因 饥饿 基因表达 |
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| 基金项目:国家高技术研究发展规划(“863”计划)项目 (2011AA100401)；国家水产种质资源平台项目；中央级公益性科研院所基本科研业务费专项 |
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| Cloning and expression analysis of NPY gene in yellow catfish，Pelteobagrus fulvidraco |
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| Abstract: |
| 【Objective】The yellow catfish (Pelteobagrus fulvidraco)NPY gene was cloned to analyze its expression pattern in adult tissues and the change of expression in brain tissue in starvation,and to investigate its physiological function in food intake.【Method】RT-PCR and RACE method were used to clone NPY gene.Bioinformatic analysis of NPY amino acid was conducted.The expression in adult tissues (brain,heart,liver,kidney,spleen,stomach,muscle,gill,and gonad) and the changes of expression in brain tissue at different starvation times (0,24,48,72,96,120,144,and 168 h) and at different times after re-feeding (1,3 and 5 h) were determined using Semi-quantitativeNPY and real-time quantitative PCR.【Result】The full-length of cDNA was 772 bp.The open reading frame was consisted of 282 bp encoding 93 amino acids.Alignment analysis showed that the amino acid sequence of Pelteobagrus fulvidraco NPY gene was 97%,87%,72%,72% and 70% identical to that of darkbarbel catfish (Pelteobagrus vachelli),channel catfish (Ictalurus punctatus),jian carp(Cyprinus carpio jian),Chinese sucker (i<>Myxocyprinus asiaticus) and barbodes sinensis(Spinibarbus sinensis),respectively.The NPY gene expression was detected in brain,spleen,liver,gill and gonad,rather than heart,muscle,stomach and kidney.The expression level in brain was much higher than in other tissues (P＜0.01),and it increased as the increase of starvation time and returned to normal level after re-feedi |
| Key words: Pelteobagrus fulvidraco NPY gene deprivation gene expression analysis |
