向日葵V-ATPasea3亚基基因的克隆及表达分析

发布时间：2018-04-24 14:43

本文选题：向日葵 + V-ATPase　；参考：《中国生物工程杂志》2017年05期

【摘要】：采用RACE技术,从向日葵P50中克隆V-ATPase a3亚基基因c DNA全长,并进行生物信息学分析;利用实时荧光定量PCR分析不同浓度、不同时间的Na Cl、ABA和PEG模拟干旱胁迫条件下V-ATPase a3亚基基因的表达特征,以及相同胁迫条件下该基因在向日葵不同器官的表达特征。序列分析表明,该基因c DNA全长2 873bp,含5'-UTR 109bp、3'-UTR 295bp及编码区2 469bp,编码822个氨基酸,其编码蛋白质的理论分子质量为204.55k Da,等电点为6.29,Gen Bank登录号为KU315054。该基因编码的蛋白质为疏水性的跨膜蛋白,亚细胞定位预测其在质膜上。向日葵V-ATPase a3亚基与已报道的10种植物的V-ATPase a3亚基的同源蛋白有高度相似的保守区域,在进化上与朝鲜蓟的亲缘关系最近。实时荧光定量PCR结果表明,向日葵受到Na Cl、ABA和PEG模拟干旱三种非生物胁迫后,V-ATPase a3亚基基因均上调表达,但表达模式不同,不同器官存在特异性表达差异。研究认为,V-ATPase a3亚基基因响应了向日葵非生物胁迫的应答,为加强对V-ATPase基因的利用奠定基础。
[Abstract]:The full length of V-ATPase a3 subunit gene c DNA was cloned from sunflower P50 by RACE technique and analyzed by bioinformatics. The expression characteristics of V-ATPase a3 subunit gene under simulated drought stress at different time and in different organs of sunflower under the same stress condition were studied. Sequence analysis showed that the full length of c DNA was 2873 BP, which contained 5'-UTR 109bpf3 + -UTR 295bp and coding region 2469 BP, encoding 822 amino acids. The theoretical molecular weight of the encoding protein was 204.55k, and the isoelectric point was 6.29gGen Bank accession number was KU315054. The protein encoded by the gene is a hydrophobic transmembrane protein, which is predicted by subcellular localization on the plasma membrane. The V-ATPase a3 subunit of sunflower has a highly conserved region with the homologous protein of V-ATPase a3 subunit of 10 reported plants, and has the closest phylogenetic relationship with artichoke. The results of real-time fluorescence quantitative PCR showed that the gene expression of V-ATPase a3 subunit was up-regulated by NaCl-ABA and PEG under abiotic stress, but the expression pattern was different and the specific expression was different in different organs. It was suggested that the subunit gene of V-ATPase A3 was responsive to abiotic stress of sunflower, which laid a foundation for enhancing the utilization of V-ATPase gene.
【作者单位】：内蒙古农业大学农学院;内蒙古农牧业科学院;
【基金】：内蒙古农牧业创新基金资助项目(2013CXJJN14)
【分类号】：S565.5

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