miR-125a/125b基因家族进化分析及其在猪睾丸组织中的表达变化

Molecular Evolution Analysis of miRNA-125a/125b Gene Family and Its Expression in Testicular Tissue of Pig

采用生物学信息方法在miRBase中搜索动物miR-125a/125b基因家族序列,利用Ensembl数据库信息确定miR-125a/125b在基因组中的位置,采用MEGA 5.0构建miR-125a/125b家族的系统进化树,并利用qRT-PCR检测miR-125a和miR-125b在沙子岭猪睾丸组织不同发育时期(D1、D30、D60、D90、D120、D150、D180)的表达变化。结果在miRBase数据库检索29个物种共58条miR-125家族序列,分布广泛,且有高度的保守性,除一个位于内含子区,两个位于外显子区,其余miR-125a/125b位于基因间隔区,在进化过程中,同种物种的进化方向具有高度的一致性,进化速度也有所不同。qRT-PCR检测结果显示,miR-125a/125b在沙子岭猪睾丸组织中不同发育时期的具有相同的趋势,即出生后其表达水平缓慢升高,保育期开始降低,性成熟期又开始呈现升高趋势,随着体成熟表达量开始降低。本研究结果为进一步探明miR-125a/125b基因家族在猪睾丸组织中的功能及作用机制奠定基础。

The miR-125a/125b gene family was searched in miRBase by using biological information method.The position of miR-125a/125b in the genome was determined based on Ensembl database information.The phylogenetic tree of miR-125a/125b gene family was constructed by using MEGA 5.0,and the expression of miR-125a and miR-125b in the testis of Shaziling pigs at different developmental stages(Dayl,30,60,90,120,150 and 180)was detected by qRT-PCR.The results showed that in the miRBase data,a total of 58 miR-125 gene family sequences were retrieved from 29 species,which were widely distributed and highly conserved.Except for one in the intron region and two in the exon region,all other miR-125a/125b genes were located in the intergenic region.In the process of evolution,the evolutionary direction of the gene was highly consistent with that of the same species,and the evolution rate was different.The result of qRT-PCR showed that miR-125a/125b had the same trend at different developmental stages in the testis of sand-ridge pigs,that was,the expression level of miR-125a/125b increased slowly after birth,decreased after weaning,and rised after the sexual maturity,and then declined as body maturation.Our results laid a foundation for further elucidating the function and mechanism of miR-125a/125b gene family in porcine testis.