本文選題：鹽穗木 + miRa��； 參考：《西北植物學報》2017年06期

【摘要】：以鹽生植物鹽穗木為實驗材料,從前期構建的高鹽脅迫下鹽穗木根的小RNA文庫中候選了差異表達的miR166a,利用生物信息學從鹽穗木轉錄組數據中預測其靶基因;采用5′RLM-RACE技術鑒定鹽穗木miR166a對預測靶基因ATHB8-like的靶向性;通過PCR和RACE技術克隆鹽穗木miR166a前體和預測靶基因ATHB8-like全長基因序列,并進行相應的生物信息學分析。結果顯示:鹽穗木miR166a預測的靶基因為ATHB8-like;通過實驗鑒定確實存在靶向切割,具體的切割位點位于miR166a成熟體的14~15堿基之間;miR166a成熟體序列在不同植物中高度保守;克隆獲得的鹽穗木miR166a前體可折疊成完整的頸環(huán)結構,符合miRNA的前體特征,候選植物miR166a前體在進化上沒有表現出保守性;預測的靶基因ATHB8-like cDNA全長為2 786bp,開放閱讀框為2 526bp,編碼841個氨基酸,ATHB8-like具有一個HD-ZIPⅢ結構域,在進化上具有保守性。該研究結果為進一步開展鹽穗木miR166a和ATHB8-like的生物學功能奠定了基礎。
[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，

本文編號：2034664