以奶牛乳腺组织为材料,采用3'RACE技术克隆到奶牛IRS1基因3'UTR的全长,进一步对测得的序列进行了验证,同时利用TargetScan 7.0和PicTar软件预测可能结合的microRNA,并对多个软件预测结合的microRNA,进行最小结合自由能分析.结果表明,成功克隆了1 327 bp的牛IRS1 3'UTR全长,与NCBI上的预测序列一致率达到99％;miR-128、miR-96、miR-27a为2个软件共同预测到的结合microRNA;3个结合位点的最小自由能分别为-89.58、-87.91、-100.88 kJ·mol-,表明牛IRS1基因表达可能受到这3个microRNA的调控.%Using bovine mammary tissue as material,bovine IRS1 gene 3'UTR full length was cloned by 3'RACE technology.Measured sequence was further validated.At the same time,TargetScan 7.0 and PicTar sofeware were used to predict the possible binding microRNA.Minimal binding free energy of the multiple software conjunct predicting microRNAs was analyzed.The results showed that the 1327bp 3'UTR full length of bovine IRS1 was successfully cloned.The detected sequence was 99％ consistent with the predicted sequences in NCBI.MiR-128,miR-96 and miR-27a were predicted by two authoritative software together.The minimum free energy of three binding sites was-89.58,-87.91 and-100.88 kJ · mol-1,respectively.These results suggested that the bovine IRS1 gene expression might be regulated by the three microRNAs.
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