山羊CMKLR1基因的克隆、表达及其与肌内脂肪的相关性研究  

Cloning and Expression of CMKLR1 Gene and Its Correlation with Intramuscular Fat in Goat(Capra hirus)

作　　者：廖红海 林亚秋[3] 朱江江[1,3] 王永[1,3] 

机构地区：[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]贵州省播州区泮水镇农业服务中心,遵义563109 [3]西南民族大学生命科学与技术学院,成都610041

出　　处：《中国畜牧兽医》2017年第6期1580-1587,共8页China Animal Husbandry & Veterinary Medicine

基　　金：国家自然科学基金(31672395;31601921);四川省"十三五"畜禽育种攻关项目(2016NZ0099-36);四川省科技创新产业链示范工程重大项目(2014NZ0003)

摘　　要：试验旨在克隆山羊CMKLR1基因序列并进行生物信息学分析,研究其表达与肌内脂肪(IMF)沉积的相关性。采集山羊皮下脂肪组织,应用RT-PCR克隆山羊CMKLR1基因序列;以GAPDH(GenBank登录号:AJ431207.1)为内参基因,实时荧光定量PCR检测其在山羊不同组织中的表达量及在肌肉组织中的时序表达谱(以肝脏表达量为基准);测定肌肉组织中的IMF含量,分析其与CMKLR1mRNA表达量的相关性。结果显示,克隆获得了山羊CMKLR1基因,1 089bp的CDS区,GenBank登录号:KT165374。实时荧光定量PCR分析显示,CMKLR1基因在24月龄山羊心脏、肝脏、脾脏、肺脏、肾脏、脂肪、背最长肌、股二头肌和臂三头肌中均存在不同程度的表达,其中在肺脏中表达量显著高于其他组织(P<0.05);CMKLR1基因在1~3和24月龄的山羊背最长肌、臂三头肌和股二头肌中表达趋势相同,均是在股二头肌中表达量最高,而8~10月龄则是在臂三头肌中表达量最高。山羊IMF含量以24月龄背最长肌含量最高。山羊背最长肌、股二头肌和臂三头肌中CMKLR1mRNA表达与IMF含量相关性不显著(P>0.05)。CMKLR1基因不参与山羊IMF沉积作用,试验结果为进一步研究CMKLR1基因奠定理论基础。In order to enrich basic date in goat （Capra hirus） CMKLR1 gene and investigate the correlation between CMKLR1 gene expression and intramuscular fat （IMF） content in muscles, the CMKLR1 gene was cloned from the goat of subcutaneous adipose tissue by RT-PCR, charac- terized by bioinformatics methods. The expression profiles of CMKLR1 gene of goat in various tissues were constructed liver was benchmark, the GAPDH （GenBank accession No.： AJ431207. 1） as a reference gene. Then, the correlation between CMKLR1 mRNA expression and IMF content in muscles was analyzed. The results showed that the CMKLR1 gene CDS ofgoat （GenBank accession No.. KT165374） was 1 089 bp. Real-time PCR indicated that CMKLR1 was widely expressed in various tissues, including heart, liver, spleen, lung, kid- ney, adipose, longissimus dorsi, biceps femoris and triceps brachii, and was the highest in lung （P〈0.05）. Similar expression variation trend of CMKLR1 was observed between the muscles from 1 to 3 and 24 months old goat, and was the highest in biceps femoris. Reversely, in 8 to 10 months old CMKLR1 was the highest in triceps brachii. The IMF of longissimus dorsi from 24 months old was the highest. Correlation analysis demonstrated that different correlations were observed between expression of CMKLR1 mRNA and IMF content in longissimus dorsi, biceps femoris, and triceps brachii in goat. The CMKLR1 gene did not participate in the depo- sition of IMF in goats. The research built the theoretical basis for further studies about the CMKLR1 gene.

关 键 词：山羊 CMKLR1基因 克隆 表达谱 肌内脂肪(IMF) 

分 类 号：S827[农业科学—畜牧学]