藏鸡肌肉组织中3条新发现miRNA序列的克隆及靶基因预测

Cloning and prediction of target genes of 3 newly discovered miRNAs in Tibetan chicken

旨在克隆藏鸡不同发育阶段肌肉组织中新发现的差异表达的3条miRNA(miR-13_529,miR-20_1207,miR-24_1302)序列,并对它们的靶基因进行预测,为3条miRNA序列功能的研究奠定基础.随机选择150日龄健康藏鸡,屠宰并采集藏鸡肌肉组织,提取组织总RNA,利用Stem-loop RT-PCR法克隆获得藏鸡3条新发现miRNA序列,利用4个生物信息学预测软件预测3条miRNA序列的靶基因.结果表明,克隆获得miR-13_529序列23bp,miR-20_1207序列21bp,miR-24_1302序列22bp.靶基因预测结果显示,分别能与3条miRNA序列特异性结合的2个、10个和1个靶基因均与肌肉生长发育相关.这些结果表明3条新发现的miRNA序列可能在藏鸡肌肉发育的过程中具有重要的调控作用.

The aim of the study was to clone 3 differentially expressed miRNAs（ miR-13_529,miR-20_1207,miR-24_1302） at different developmental stages of muscle tissue in Tibetan chicken,and to predict the target genes of 3 miRNAs. This study might provide basis for its functional research on regulating the growth and development of muscle in Tibetan chicken. A 150-day-old healthy Tibetan chicken was selected after being slaughtered,and the muscle tissue was collected for RNA extraction. The sequences of 3 newly discovered miRNAs were cloned by Stem-loop RT-PCR,and the target genes of the 3 miRNAs were predicted by 4 bioinformatics softwares. The result showed that the cloned sequences of 3 miRNAs were 23 bp,21 bp and22 bp respectively. The predicted 2,10 and 1 target genes of 3 miRNAs were related to the growth and development of muscle.These results suggest that 3 newly discovered miRNAs may play important roles in regulating the development of muscle in Tibetanchicken.