巴什拜羊miR-486多态性及其表达规律研究  

Polymorphism of miR-486 and Its Expression Pattern in Bashby Sheep(Ovis aries)

作　　者：张伟[1,2] 王世银[1] 石国庆[2] 邓双义[1] 刘晓娜[1] 杨力伟[1] ZHANG Wei;WANG Shi-Yin;SHI Guo-Qing;DENG Shuang-Yi;LIU Xiao-Na;YANG Li-Wei(Xinjiang Agricultural Professional Technological College,Changji 831100,China;Institute of Animal Husbandry and Veterinary Medicine,Xinjiang Academy of Agricultural and Reclamation Sciences,Shihezi 832000,China)

机构地区：[1]新疆农业职业技术学院,昌吉831100 [2]新疆农垦科学院畜牧兽医研究所,石河子832000

出　　处：《农业生物技术学报》2020年第1期92-100,共9页Journal of Agricultural Biotechnology

基　　金：新疆维吾尔自治区自然科学基金面上项目(2017D01A51)

摘　　要：miR-486在动物肌肉组织中高表达,在成肌细胞的生长发育中发挥着关键作用。为了阐明miR-486在巴什拜羊(Ovis aries)中的多态性及其表达规律,本研究采用stem-loop qRT-PCR对巴什拜羊胎儿期100 d不同组织,以及胎儿期和出生后共计8个不同发育阶段骨骼肌中miR-486的表达情况进行了定量分析;采用PCR-单链构象多态性(PCR-single strand conformation polymorphism,PCR-SSCP)技术检测了巴什拜羊群体中miR-486前体(precursor miR-486,pre-miR-486)序列的突变情况,并在细胞水平研究了突变对pre-miR-486序列加工成熟的影响。结果表明,miR-486在巴什拜羊100 d胎儿心肌和骨骼肌组织中高表达,在胃、肠组织中度表达,而在肝、脾、肺、肾、皮肤和脑组织中表达量相对较低;miR-486在胎儿期40~100 d的骨骼肌中表达量持续上升,到胎儿期100 d时达到峰值,然后逐渐下降,至胎儿期120 d时达到最低值,而后又逐渐上升直至出生后90 d;在巴什拜羊群体中发现pre-miR-486序列存在1处G>C突变,位于miR-486成熟序列上游第1个碱基处,在100只巴什拜羊中分别检测到GG、GC和CC 3种基因型,但91%的个体为GC基因型,进一步研究结果表明,在骨骼肌卫星细胞中突变型的CC基因型可显著提高细胞中miR-486的表达水平(P<0.05)。本研究可为进一步阐明miR-486对巴什拜羊骨骼肌发育的调控机制及其与巴什拜羊优良肉质性状的关联性提供基础资料。miR-486 is expressed in animal muscle at a high level and plays a key role in growth and development of myoblast in different tissues.To illuminate the polymorphism and expression pattern of miR-486 in Bashby sheep(Ovis aries),the stem-loop qRT-PCR was used to detect the expression levels in different tissues,and in skeletal muscle tissues of 8 fetal and postnatal periods of Bashby sheep.Then the mutation of miR-486 sequence in Bashby sheep group and its effect on cutting process of precursor miR-486(pre-miR-486)in skeletal muscle cell were also researched.The result showed that the miR-486 was expressed at a high level in myocardium and skeletal muscle of the 100 d of fetal period of Bashby sheep,moderately expressed in stomach and intestines,and lowly expressed in liver,spleen,lung,skin and brain tissues;The expression levels of miR-486 in skeletal muscle of the 40~100 d of fetal periods were increased gradually,to a peak level on the100 d of fetal periods then decreased gradually,and to a minimum level on the 120 d of fetal periods then increased continually until the 90 d of postnatal period;A G>C mutation was detected in pre-miR-486 sequence,which located at the first base of upstream of miR-486 mature sequence,the GG,GC and CC genotype could be detected in Bashby sheep group,but 91%individual of them were GC genotype,the further investigation indicated that the CC genotype could significantly up regulate the level of miR-486 in skeletal muscle cell(P<0.05).The present research could provide basic data for further uncovering the regulation mechanism of miR-486 in development of Bashby sheep skeletal muscle and relationship between its expression and excellent meat quality traits of Bashby sheep.

关 键 词：巴什拜羊 miR-486 突变 骨骼肌 卫星细胞 

分 类 号：S826.8[农业科学—畜牧学]