摘要：

bta-miR-6523a是前期史远刚课题组通过测序数据筛选出的牛脂肪沉积相关基因之一。为筛选bta-miR-6523a的靶基因,鉴定其在不同组织中的表达规律,探索其对牛脂肪沉积的可能调控机制,采用多种数据库信息对bta-miR-6523a靶基因进行预测及筛选,并探究预测的靶基因功能富集通路,最后采用qRT-PCR方法检测bta-miR-6523a在成年牛和犊牛各组织中的表达情况。结果表明,bta-miR-6523a的预测靶基因有SCD、CIDEA、LIPE等;这些预测靶基因显著富集于Ras、Rap1、肌动蛋白细胞骨架的调节以及Hippo等与脂肪细胞的生成与分化、细胞骨架的组织和维护、代谢途径相关信号通路中。qRT-PCR结果表明,牛miR-6523a在肺中的表达量最高,在肾周脂肪中的表达量最低;bta-miR-6523a在心和肝中的表达显著高于皮下脂肪、肌间脂肪、背最长肌以及肾周脂肪,皮下脂肪、肌间脂肪、背最长肌以及肾周脂肪之间差异不显著;miR-6523a在犊牛皮下脂肪、肌间脂肪和背最长肌中的表达差异显著,且显著低于成年牛组织;bta-miR-6523a在肺组织中的高表达以及在脂肪组织表达低于其他组织提示bta-miR-6523a及其靶基因生物学作用可能较为广泛。

关键词: 牛, bta-miR-6523a, 靶基因预测, 功能富集

Abstract:

bta-miR-6523a is one of the genes related to bovine fat deposition screened by the previous research Shi Yuangang's group through sequencing data.In order to screen the target gene of bta-miR-6523a,identify its expression rule in different tissues,and explore the possible regulatory mechanism of bta-miR-6523a on bovine fat deposition,this study used a variety of database information to predict and screen the target gene of bta-miR-6523a,and enriched and analyzed the predicted functional pathway of the target gene.Finally,the expression of bta-miR-6523a in some tissues of adult cattle and calves was detected by qRT-PCR.The results showed that the predicted target genes of bta-miR-6523a were SCD,CIDEA,LIPE,etc.These predicted target genes were significantly enriched in Ras,Rap1,actin cytoskeleton regulation,Hippo and other signaling pathways related to adipocyte generation and differentiation,cytoskeleton organization and maintenance,and metabolic pathways.qRT-PCR results showed that the expression level of bovine miR-6523a was the highest in lung and the lowest in perirenal fat.The expression of bta-miR-6523a in heart and liver was significantly higher than that in subcutaneous fat,intermuscular fat,longissimus dorsi muscle and perirenal fat,but there was no significant difference among subcutaneous fat,intermuscular fat,longissimus dorsi muscle and perirenal fat.The expression of miR-6523a in subcutaneous fat,intermuscular fat and longissimus dorsi muscle of calves was significantly different,and significantly lower than that in adult bovine tissues.The high expression of bta-miR-6523a in lung tissue and lower expression in adipose tissue suggest that the biological effects of bta-miR-6523a and its target genes may be extensive.

Key words: Cattle, bta-miR-6523a, Target gene prediction, Functional enrichment