白菜型油菜MPK12基因克隆及表达分析
发布时间：2019-01-20 17:06
【摘要】：【目的】克隆白菜型油菜‘陇油6号’MPK12基因的全长cDNA序列,研究其组织表达特异性,分析MPK12基因在低温、盐、ABA和H_2O_2处理下的表达情况,以阐明MPK12基因在油菜中的生物学功能。【方法】利用RACE技术克隆MPK12基因cDNA全长,并对其全长基因进行生物信息学分析;构建系统发育树,研究其与相似序列的同源性;利用实时荧光定量PCR方法,分析MPK12基因的组织表达特异性以及在低温、盐、ABA和H_2O_2逆境胁迫下的表达情况。【结果】油菜MPK12基因cDNA全长1 395bp,包括5′-UTR 69bp,3′-UTR 207bp,开放阅读框1 119bp,编码372个氨基酸,预测蛋白质分子量42.6ku,理论等电点为7.9,二级结构主要包括α-螺旋和不规则卷曲。多序列比对和系统进化分析表明,油菜MPK12与拟南芥AtMPK12具有很高的同源性,为90.7%。实时荧光定量PCR结果显示,MPK12基因在油菜根、茎、叶、芽和种子中均有表达,没有组织特异性;同时,该基因的表达受低温、盐、ABA和H_2O_2胁迫诱导。【结论】克隆得到油菜MPK12基因,其在油菜适应逆境胁迫过程中发挥作用。
[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 伪 -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.
【作者单位】： 西北师范大学生命科学学院;
【基金】：国家自然科学基金项目(31460099,31160089) 甘肃省自然科学基金项目(1208RJZA268)
【分类号】：S565.4
，
本文编号：2412216
[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 伪 -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.
【作者单位】： 西北师范大学生命科学学院;
【基金】：国家自然科学基金项目(31460099,31160089) 甘肃省自然科学基金项目(1208RJZA268)
【分类号】：S565.4
，
本文编号：2412216
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