【摘要】：【目的】克隆白菜型油菜‘隴油6號’MPK12基因的全長cDNA序列,研究其組織表達(dá)特異性,分析MPK12基因在低溫、鹽、ABA和H_2O_2處理下的表達(dá)情況,以闡明MPK12基因在油菜中的生物學(xué)功能。【方法】利用RACE技術(shù)克隆MPK12基因cDNA全長,并對其全長基因進(jìn)行生物信息學(xué)分析;構(gòu)建系統(tǒng)發(fā)育樹,研究其與相似序列的同源性;利用實(shí)時(shí)熒光定量PCR方法,分析MPK12基因的組織表達(dá)特異性以及在低溫、鹽、ABA和H_2O_2逆境脅迫下的表達(dá)情況�！窘Y(jié)果】油菜MPK12基因cDNA全長1 395bp,包括5′-UTR 69bp,3′-UTR 207bp,開放閱讀框1 119bp,編碼372個(gè)氨基酸,預(yù)測蛋白質(zhì)分子量42.6ku,理論等電點(diǎn)為7.9,二級結(jié)構(gòu)主要包括α-螺旋和不規(guī)則卷曲。多序列比對和系統(tǒng)進(jìn)化分析表明,油菜MPK12與擬南芥AtMPK12具有很高的同源性,為90.7%。實(shí)時(shí)熒光定量PCR結(jié)果顯示,MPK12基因在油菜根、莖、葉、芽和種子中均有表達(dá),沒有組織特異性;同時(shí),該基因的表達(dá)受低溫、鹽、ABA和H_2O_2脅迫誘導(dǎo)�！窘Y(jié)論】克隆得到油菜MPK12基因,其在油菜適應(yīng)逆境脅迫過程中發(fā)揮作用。
[Abstract]:[objective] to clone the full-length cDNA sequence of MPK12 gene of Brassica campestris' Longyou 6', to study its tissue expression specificity, and to analyze the expression of MPK12 gene under low temperature, salt, ABA and H_2O_2 treatments. In order to elucidate the biological function of MPK12 gene in rapeseed. [methods] the full length of MPK12 gene cDNA was cloned by RACE and its full-length gene was analyzed by bioinformatics. The phylogenetic tree was constructed to study its homology with similar sequences. The tissue expression specificity of MPK12 gene and its expression under stress of low temperature, salt, ABA and H_2O_2 were analyzed by real-time fluorescence quantitative PCR. [results] the length of cDNA of MPK12 gene in rapeseed was 139.5 BP, including 5'-UTR 69bp. 3'-UTR 207 BP, open reading frame 1119 BP, encoding 372 amino acids, predicted protein molecular weight 42.6 ku.The theoretical isoelectric point is 7.9. The secondary structure mainly includes 偽 -helix and irregular curl. Multiple sequence alignment and phylogenetic analysis showed that rape MPK12 had high homology with Arabidopsis thaliana AtMPK12 (90.7). The results of real-time fluorescence quantitative PCR showed that MPK12 gene was expressed in root, stem, leaf, bud and seed of rape without tissue specificity. At the same time, the expression of this gene was induced by low temperature, salt, ABA and H_2O_2 stress. [conclusion] the MPK12 gene was cloned into rapeseed, which plays an important role in the process of rapeseed adaptation to stress.
【作者單位】： 西北師范大學(xué)生命科學(xué)學(xué)院;
【基金】：國家自然科學(xué)基金項(xiàng)目(31460099,31160089) 甘肅省自然科學(xué)基金項(xiàng)目(1208RJZA268)
【分類號】：S565.4
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本文編號：2412216