山羊FGF21基因克隆及其在肌内脂肪细胞中的表达模式研究  

Cloning of Goat FGF21 Gene and Its Expression Pattern in Intramuscular Adipocyte

作　　者：李倩[1,2] 林亚秋[1] 朱江江[1,2] 林森[1,2] 张小玉[1] 王永[1,2] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041 [2]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

出　　处：《畜牧兽医学报》2017年第1期31-38,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：四川省"十三五"畜禽育种攻关项目(2016NYZ0045);四川省科技创新产业链示范工程重大项目(2014NZ0003);中央高校基本科研业务费专项资金项目(2016NZYQN36)

摘　　要：本研究旨在获得山羊FGF21基因序列,阐明其组织表达特性,同时检测FGF21基因与FGF受体(FGFR)在山羊肌内前体脂肪细胞分化过程中的表达水平。以简州大耳羊为试验动物,采用II型胶原酶消化获得山羊原代肌内前体脂肪细胞,采用RT-PCR技术克隆山羊FGF21基因,利用荧光定量PCR(qPCR)技术检测FGF21基因在成年山羊不同组织中的表达特性及FGF21与FGF受体在山羊肌内前体脂肪细胞分化过程中的表达差异。结果,克隆获得山羊FGF21基因(GenBank登录号:KT288195)全长序列666bp,其中开放阅读框630bp,编码209个氨基酸。FGF21mRNA在山羊心、肝、脾、肺、肾、脂肪和背最长肌中均检测到表达,且在肝和脂肪中存在较高水平表达。FGF21基因与FGFR1、FGFR2表达模式相同,均是在诱导分化第2天的肌内脂肪细胞中表达水平最高,极显著高于其他天数(P<0.01)。FGF21基因在山羊脂肪组织中存在较高水平的表达,并且在诱导分化的肌内前体脂肪细胞成脂分化第2天表达水平最高,推测其可能通过受体FGFR1或FGFR2在山羊肌内前体脂肪细胞分化早期发挥调控作用,此研究结果为进一步揭示FGF21基因在山羊肌内脂肪沉积中的作用机制提供了数据支持。This study was conducted to obtain the sequence of goat FGF21 gene,elucidate the expression characteristics of FGF21 gene in different tissues,and detect the expression of FGF21 and FGFRduring intramuscular preadipocyte differentiation.Jianzhou Big-eared goat was used as laboratory animal in this study.The goat intramuscular preadipocytes were obtained using collagenase II.RT-PCR was used to clone FGF21 gene.Real-time quantitative PCR（qPCR）was performed to detect the expression of FGF21 in different tissues and the expression levels of FGF21 and its receptors during goat intramuscular preadipocyte differentiation.The full-length sequence of goat FGF21 gene was 666 bp including a complete 630 bp open reading frame（ORF）,encoding209 amino acids（Accession No.：KT288195）.FGF21 gene was expressed in all examined tissues including heart,liver,spleen,lung,kidney,adipose tissue and longissimus dorsi,and enriched in liver and adipose tissue.During the differentiation of goat intramuscular preadipocyte,the ex-pression pattern of FGF21 gene was the same as that of FGFR1 and FGFR2,peaked at 2nd day,extremely higher than other days（P〈0.01）.The FGF21 mRNA was enriched in adipose tissue and peaked at 2nd day during goat intramuscular preadipocyte differentiation,indicating it might be a regulator in the early differentiation progress via FGFR1 or FGFR2.These results provide data for further elucidating the molecular mechanism of FGF21 gene in regulating goat IMF deposition.

关 键 词：FGF21 FGFR 山羊 克隆 表达 肌内前体脂肪细胞 

分 类 号：S827[农业科学—畜牧学] S813.1[农业科学—畜牧兽医]