盐穗木miR166a前体和预测靶基因ATHB8-like的克隆及二者靶向关系的鉴定

发布时间：2018-06-18 07:22

  本文选题：盐穗木 + miRa　； 参考：《西北植物学报》2017年06期

【摘要】：以盐生植物盐穗木为实验材料,从前期构建的高盐胁迫下盐穗木根的小RNA文库中候选了差异表达的miR166a,利用生物信息学从盐穗木转录组数据中预测其靶基因;采用5′RLM-RACE技术鉴定盐穗木miR166a对预测靶基因ATHB8-like的靶向性;通过PCR和RACE技术克隆盐穗木miR166a前体和预测靶基因ATHB8-like全长基因序列,并进行相应的生物信息学分析。结果显示:盐穗木miR166a预测的靶基因为ATHB8-like;通过实验鉴定确实存在靶向切割,具体的切割位点位于miR166a成熟体的14~15碱基之间;miR166a成熟体序列在不同植物中高度保守;克隆获得的盐穗木miR166a前体可折叠成完整的颈环结构,符合miRNA的前体特征,候选植物miR166a前体在进化上没有表现出保守性;预测的靶基因ATHB8-like cDNA全长为2 786bp,开放阅读框为2 526bp,编码841个氨基酸,ATHB8-like具有一个HD-ZIPⅢ结构域,在进化上具有保守性。该研究结果为进一步开展盐穗木miR166a和ATHB8-like的生物学功能奠定了基础。
[Abstract]:Using halophyte halophytes as experimental materials, the differentially expressed miR166a was selected from the small RNA library of salt panicle root constructed under high salt stress, and its target gene was predicted from the transcription data of salt panicle by bioinformatics. The target of the predicted target gene ATHB8-like was identified by 5RLM-RACE, and the full-length sequence of the precursor and the predicted target gene ATHB8-like were cloned by PCR and race, and the corresponding bioinformatics analysis was carried out. The results showed that the target base predicted by MiR166a was ATHB8-like, and the specific cleavage site was located between 1415 bases of miR166a mature plant, which was highly conserved in different plants. The cloned miR166a precursor can be folded into a complete cervical ring structure, which accords with the characteristics of miRNA precursor, but the candidate plant miR166a precursor does not show conservation in evolution. The predicted target gene ATHB8-like cDNA is 2786 BP, and the open reading frame is 2526 BP, encoding 841 amino acids, ATHB8-like has a HD-ZIP 鈪，

本文编号：2034664