摘要
本文旨在克隆山羊GPR1基因,对其进行序列分析,并研究组织表达。以简州大耳羊为试验材料,应用RT-PCR方法克隆山羊GPR1基因并进行生物信息学分析,利用实时荧光定量PCR构建组织表达谱。结果表明:山羊GPR1基因编码区长度为987 bp(Gen Bank登录号:KT347601),编码蛋白为稳定疏水蛋白;存在7个跨膜结构域,在69~304氨基酸序列属于G蛋白偶联受体家族保守结构域(Pfam:7tm-1);荧光定量PCR分析表明,GPR1在24月龄山羊心、肝、脾、肺、肾、脂肪、背最长肌、股二头肌和臂三头肌中均有不同程度的表达,其中肺脏中表达量最高且极显著高于其他组织(P〈0.01),脂肪、脾、肝和心次之,股二头肌中表达量最低;GPR1基因在1~3月龄与24月龄表达变化趋势相同,由背最长肌、股二头肌和臂三头肌呈现逐渐上升的趋势,而8~10月龄则呈现相反的变化趋势。该实验结果为进一步研究GPR基因提供理论基础。
The aim of this study was to clone and analyze the GPR1 gene sequence,and to characterize the expression profile in various tissues of Jianzhou Da’er Goat.The GPR1 gene was cloned by RT-PCR,and analyzed using bioinformatics methods.The tissue expression was analyzed using the fluorescence quantitative PCR.The resultsshowed that the CDS of GPR1 gene was 987 bp(Gen Bank accession No.KT347601),encoding a stable and unhydrophilic protein.The GPR1 protein contained 7 protein transmembrane regions with a conserved protein domain(Pfam:7tm-1) of G protein-coupled receptors family at 69 ~304 aa.Fluorescence quantitative PCR indicated that GPR1 was widely expressed in various tissues,including heart,liver,spleen,lung,kidney,adipose,longissimus dorsi muscle,biceps femoris muscle and triceps brachii muscle.The m RNA expression level of the GPR1 gene was highest in lung(P〈0.01),followed by adipose,spleen,live and heart,and lowest in biceps femoris muscle.Similar expression variation trend of GPR1 was observed between the muscles from 1 ~3 and 24 months goat,highest in longissimus dorsi muscle and lowest in triceps brachii muscle,reversedly in 8 ~10 months.The research built the theoretical basis for further studies about the GPR1 gene.
出处
《中国畜牧杂志》
CAS
北大核心
2016年第11期5-10,共6页
Chinese Journal of Animal Science
基金
四川省"十二五"畜禽育种攻关项目(2011NZ0099-36)
四川省科技创新产业链示范工程重大项目(2014NZ0003)
四川省教育厅项目(15ZA0385)