猪microRNA-17-92基因簇分析

Analysis on porcine microRNA-17-92 cluster

为了分析猪microRNA-17-92基因簇的结构和表达特性,试验采用生物信息学方法分析其序列保守性、PCR产物直接测序的方法检测其在民猪、荷包猪和大白猪中的多态性,并利用荧光定量PCR技术构建其原初转录本和成熟microRNA-18a的组织表达谱。结果表明：microRNA-17-92基因簇各成熟miRNA在猪、人、小鼠、鸡和非洲爪蟾等物种中高度保守;在microRNA-17-92基因簇3＇侧翼区发现一个SNP位点,为中度多态,各等位基因在民猪、荷包猪和大白猪群体中具有相似的频率;microRNA-17-92的原初转录本和成熟microRNA-18a普遍表达于所检测的组织中,但表达模式存在差异。说明猪microRNA-17-92基因簇各成熟miRNA受到严格的表达调控。

The aim of the present study was to analyze the structure and expression characteristics of porcine microRNA-17-92 cluster. The sequence conservatism was identified by using bioinformatic methods. The polymorphisms of the genes in Min pig,Hebao pig and Yorkshire were revealed by direct sequencing of PCR products. The primary transcripts and expression profiles of mature microRNA-18 a were constructed by fluorescence quantitative PCR. The results showed that the each mature miRNA of the microRNA-17-92 gene cluster was highly conserved in pig,human,mouse,chicken and Xenopus species. A SNP locus was found in the 3＇ flanking region of the microRNA-17-92 gene cluster,which was moderately polymorphic. The alleles had similar frequencies in the population of Min pig,Hebao pig and Yorkshire. The primary transcripts of microRNA-17-92 and mature microRNA-18 a were generally expressed in the tested tissues,but the expression profiles were different. These data indicated that the expression of each miRNA in the clusters was strictly regulated.