兔神经肽W及其受体NPWR1基因克隆及分布定位.pdfVIP

中国农业科学 20 12,45(11):2295-2305 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2012.11.020 兔神经肽W 及其受体NPWR1 基因克隆及分布定位 孙晓璇 雷治海 杨桂红 刘新义 叶 平 （南京农业大学动物医学院，南京210095 ） 摘要：【目的】研究神经肽W （NPW）及其受体1 （NPWR1，GPR7）mRNA 在兔各器官组织的表达情况。【方法】 利用基因克隆方法克隆出兔神经肽W 及其受体的cDNA 序列片段，以GAPDH 为内参基因，采用相对定量RT-PCR 方 法检测NPW 及NPWR1 mRNA 在兔体内各组织中的表达情况，用原位杂交方法研究NPWR1 阳性细胞在兔脑的分布定 位。 【结果】克隆出兔NPW 及其受体NPWR1 的基因部分序列，NPW 基因片段长234 bp，NPWR1 基因片段长508 bp， GenBank 登录号分别为HQ596517 和HQ596518；经DNAStar 软件分析，不同物种之间NPW 及其受体基因序列的同 源性较高。NPW 及其受体在兔各组织广泛分布；NPWR1 mRNA 在脑内主要分布于海马、小脑、下丘脑、延髓等部位。 【结论】兔NPW 及其受体基因在进化上是保守的；NPW 及其受体在兔体内广泛表达，提示其参与多种生理功能的 调节。 关键词：神经肽W；神经肽W 受体；基因克隆；分布定位；兔 Gene Cloning and Distribution of Neuropeptide W and Its Receptor 1 in Rabbit SUN Xiao-xuan, LEI Zhi-hai, YANG Gui-hong, LIU Xin-yi, YE Ping (College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095) Abstract: 【Objective 】 This study was carried out to obtain sequence of NPW and NPWR1 (GPR7) gene, and reveal the expression of NPW and NPWR1 mRNA in rabbit organs. 【Method 】 The partial coding sequences of NPW and NPWR1 were obtained from rabbit brain using the approach of gene cloning. The expressions of NPW and NPWR1 mRNA in rabbit brain were studied by semi-quantitative RT-PCR and in situ hybridization. 【Result 】The length of NPW and NPWR1 was 234 bp and 508 bp, respectively. The sequences of the products were confirmed to be those of NPW and NPWR1 in rabbit and enrolled in GenBank (accession numbers: HQ596517 and HQ596518). Phylogenetic analysis showed that amino sequence of rabbit NPS precursor had high homology compared with other animals. The expression of NPW and NPWR1 mRNA was extensive in rabbit organs, with maj