本文選題：鹽穗木 + miRa�。� 參考：《西北植物學(xué)報(bào)》2017年06期

【摘要】：以鹽生植物鹽穗木為實(shí)驗(yàn)材料,從前期構(gòu)建的高鹽脅迫下鹽穗木根的小RNA文庫中候選了差異表達(dá)的miR166a,利用生物信息學(xué)從鹽穗木轉(zhuǎn)錄組數(shù)據(jù)中預(yù)測其靶基因;采用5′RLM-RACE技術(shù)鑒定鹽穗木miR166a對預(yù)測靶基因ATHB8-like的靶向性;通過PCR和RACE技術(shù)克隆鹽穗木miR166a前體和預(yù)測靶基因ATHB8-like全長基因序列,并進(jìn)行相應(yīng)的生物信息學(xué)分析。結(jié)果顯示:鹽穗木miR166a預(yù)測的靶基因?yàn)锳THB8-like;通過實(shí)驗(yàn)鑒定確實(shí)存在靶向切割,具體的切割位點(diǎn)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(jié)構(gòu),符合miRNA的前體特征,候選植物miR166a前體在進(jìn)化上沒有表現(xiàn)出保守性;預(yù)測的靶基因ATHB8-like cDNA全長為2 786bp,開放閱讀框?yàn)? 526bp,編碼841個氨基酸,ATHB8-like具有一個HD-ZIPⅢ結(jié)構(gòu)域,在進(jìn)化上具有保守性。該研究結(jié)果為進(jìn)一步開展鹽穗木miR166a和ATHB8-like的生物學(xué)功能奠定了基礎(chǔ)。
[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