山羊CC类趋化因子受体（CCR2）基因的克隆及组织表达分析

doi:10.11843/j.issn.0366-6964.2016.11.007

畜牧兽医学报 ›› 2016, Vol. 47 ›› Issue (11): 2202-2209.doi: 10.11843/j.issn.0366-6964.2016.11.007

山羊CC类趋化因子受体（CCR2）基因的克隆及组织表达分析

朱江江¹，林亚秋^2*，王永^1,2，李倩^1,2，林森^1,2

（1. 青藏高原动物遗传资源保护与利用四川省重点实验室，成都 610041；2. 西南民族大学生命科学与技术学院，成都 610041)

收稿日期:2016-03-30 出版日期:2016-11-23 发布日期:2016-11-23
通讯作者: 林亚秋，博士，研究员，硕士生导师，主要从事动物遗传育种研究，E-mail: linyq1999@163.com
作者简介:朱江江(1986-)，男，湖北钟祥人，博士，助理研究员，主要从事动物遗传育种研究，E-mail: zhujiang4656@hotmail.com
基金资助:
中央高校基本科研业务费专项资金项目 (2015NZYQN80)；四川省应用基础研究计划(2016JY0147)

Molecular Cloning and Tissue Expression of Goat Chemokine (CC motif) Receptor 2 (CCR2) Gene

ZHU Jiang-jiang¹, LIN Ya-qiu^2* , WANG Yong^1,2, LI Qian^1,2, LIN Sen^1,2

(1. Key Laboratory of Sichuan Province for Qinghai-Tibetan Plateau Animal Genetic Resource Reservation and Exploitation, Chengdu 610041, China; 2. College of Life Science and Technology, Southwest University for Nationalities, Chengdu 610041, China)

Received:2016-03-30 Online:2016-11-23 Published:2016-11-23

摘要/Abstract

摘要：

本研究旨在克隆山羊CC类趋化因子受体基因（CCR2）的CDS区序列，并检测其在山羊不同组织中mRNA和蛋白表达水平，从而为CCR2基因在山羊脂代谢中的作用研究奠定基础。选取2～3周岁的健康简州大耳羊和藏山羊羯羊各4只，屠宰后分别采集心、肝、脾、肺、肾、肌肉和皮下脂肪组织，分别提取组织总RNA及总蛋白，利用RT-PCR法克隆山羊CCR2基因序列，进行生物信息学分析，并利用实时定量PCR法和Western blot分别检测CCR2基因mRNA和蛋白的表达水平。结果显示，克隆得到CCR2基因cDNA序列1 186 bp，包含CDS全长1 100 bp，5′ UTR 2 bp 和3′ UTR 74 bp，编码370个氨基酸残基。山羊CCR2氨基酸序列与牛、猪、小鼠、人的相似性分别为97.3%、91.9%、77.0%和69.2%。山羊CCR2基因与牛具有最近的亲缘关系，其次为猪、狗和小鼠，而与人的亲缘关系较远。组织表达结果显示，CCR2在两种山羊肾和心中均具有较高的表达水平，而在皮下脂肪和肌肉中的表达水平较低。这些结果表明，CCR2可能在山羊脂质代谢过程中具有重要的调控作用。

Abstract:

The aim of the study was to clone the CDS of CCR2 gene, and to determine its expression in various tissues, which might provide basis for its functional research on regulating lipid metabolism. Each of 4 healthy, 2-3 years old castrated Jianzhou Big-eared goats and Tibetan goats were selected. After slaughter, the tissue samples from heart, liver, spleen, lung, kidney, longissimus dorsi muscle, biceps femoris muscle, triceps brachii muscle and subcutaneous fat were collected for the total protein and RNA extraction. The sequence of CCR2 gene was cloned by RT-PCR and analyzed using bioinformatics. The mRNA and protein expression of CCR2 was determined using real-time fluorescent quantitative (qPCR) and Western blot, respectively. A length of 1 186 bp cDNA of CCR2 gene was cloned, containing 1 100 bp CDS, 2 bp 5′ UTR and 74 bp 3′ UTR, encoding 370 amino acids. The goat CCR2 protein shared 97.3%, 91.9%, 77.0% and 69.2% similarity with bovine, pig, mouse and human, respectively. The goat CCR2 gene was closest associated with that of bovine, followed by pig, dog and mouse, and distant with human. The tissue expression analysis showed that expression of CCR2 was higher in kidney and heart, and lowest in subcutaneous fat and muscles in both of the goat breeds. These data indicate that CCR2 gene may play an important role in regulating lipid metabolism in goat.

中图分类号:

S827
S813.3

朱江江，林亚秋，王永，李倩，林森. 山羊CC类趋化因子受体（CCR2）基因的克隆及组织表达分析[J]. 畜牧兽医学报, 2016, 47(11): 2202-2209.

ZHU Jiang-jiang, LIN Ya-qiu, WANG Yong, LI Qian, LIN Sen. Molecular Cloning and Tissue Expression of Goat Chemokine (CC motif) Receptor 2 (CCR2) Gene[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(11): 2202-2209.

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山羊CC类趋化因子受体（CCR2）基因的克隆及组织表达分析

Molecular Cloning and Tissue Expression of Goat Chemokine (CC motif) Receptor 2 (CCR2) Gene

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