星斑川鲽神经肽Y基因cDNA克隆与表达特性研究  

cDNA Cloning and Expression Pattern of Neuropeptide Y cDNA in Platichthys stellatus

作　　者：郭湘云[1,2] 李华[1] 郑风荣[2] 王波[2] 徐宗军[2] 丁倩倩[2] 赵盟[2] 李青[2] 

机构地区：[1]大连海洋大学水产与生命科学学院,辽宁大连116023 [2]国家海洋局第一海洋研究所,山东青岛266000

出　　处：《海洋科学进展》2015年第3期352-360,共9页Advances in Marine Science

基　　金：国家高技术研究发展计划项目--星斑川鲽养殖技术研究(2012AA10413);海洋公益性行业科研专项--环境工程生态优化;技术集成应用效果评估(201305005)

摘　　要：神经肽Y(Neuropeptide Y,NPY)是一种单链多肽,被认为是促进脊椎动物摄食的一个重要诱导因子。以星斑川鲽为研究对象,通过SMART-RACE技术得到了星斑川鲽NPY基因的cDNA序列,其长度为559bp,包含13bp的5′非编码区、210bp的开放阅读框和334bp的3′非编码区,该基因编码69个氨基酸,理论分子量为7.98kDa,等电点为5.24。与已知物种神经肽Y进行同源性比对,得知与鲽科黄盖鲽属鱼类的NPY属于一支。利用实时荧光定量PCR技术检测了神经肽Y在星斑川鲽不同组织中的表达,以及饥饿对该基因在脑组织中的表达影响,Real-time PCR结果显示在性腺、肠、肾、肝、心脏、脑组织中NPY mRNA均有表达,但在表达量上有明显差异,心脏和肾组织相对表达量较少;且随着饥饿时间的增长,NPY在脑组织中表达量逐渐增加,饥饿72h的相对表达量高于饥饿24h的,但差异不显著(P>0.05)。本研究可为星斑川鲽的繁殖发育以及遗传育种提供理论基础。Neuropeptide Y（NPY）is a single chain polypeptide,and is considered as an important factor to promote feeding of the invertebrates.In the present study,full length cDNA sequence of the Neuropeptide Y（NPY）gene was cloned from brain tissue of Platichthys stellatus by using 3′RACE and 5′RACE,and the sequence and structural analysis of the NPY gene were investigated by using bioinformatics method.The results showed that the NPY cDNA sequence consisted of 559 bp nucleotides,including 13 bp at 5′untranslation region,334 bp at 3′untranslation region and an open reading frame with 210 bp nucleotides,which could encode 69 amino acids.And the theoretical molecular weight was 7.98 kDa,pI was 5.24.Comparison of the amino acid sequences and phylogenetic analysis showed that the Platichthys stellatus NPY had the close relationship with Limanda,which was highest similarity with Pseudopleuronectes americanus（100%）.Meantime,the results suggesteded that the level of NPY mRNA was higher in brain,gonads liver,intestines and lower in heart and kidney by Real-time Quantitative PCR technique,and with the increase of starvation（0,24,72h）,the NPY mRNA expression in brain was gradually rising,but did not exhibit a significant difference（P〉0.05）.This research provided a theoretical basis for reproductive development and genetic breeding of Platichthys stellatus.

关 键 词：星斑川鲽 神经肽Y CDNA克隆 序列分析 表达 

分 类 号：Q75[生物学—分子生物学]