白骨壤重金属耐受性相关基因AmNramp3的克隆与分析

发布时间：2018-03-21 04:41

本文选题：白骨壤　切入点：重金属耐受性　出处：《厦门大学学报(自然科学版)》2017年03期 　论文类型：期刊论文

【摘要】：白骨壤(Avicennia marina(Forsk.)Vierh.)作为红树林的先锋植物,广泛分布于我国南海沿岸,是一种研究红树植物重金属污染物耐受性分子机制很好的材料.以抑制消减杂交获得的白骨壤重金属耐受性相关基因AmNramp3片段为基础,设计基因特异性引物,利用cDNA末端快速扩增(RACE)技术克隆了其全长,长度为1 958bp;通过实时荧光定量PCR(qRT-PCR)检测了AmNramp3在重金属镉胁迫时白骨壤叶、茎和根中的表达情况,其表达量在叶中高于根和茎中,但在茎和根中有快速的响应.利用生物信息学方法预测并分析了该基因的开放阅读框,位置位于1~1 527bp,编码508个氨基酸;进而预测并分析了AmNramp3蛋白的结构,发现其氨基酸序列中不含有信号肽,说明其不是分泌蛋白;但其具有很复杂的跨膜结构,是一种跨膜蛋白.综上,AmNramp3基因编码一种跨膜蛋白,在重金属镉胁迫下表达量快速变化,可能与镉的跨膜运输密切相关,这为后续分析该基因发挥功能的分子机制奠定了基础.
[Abstract]:As a pioneer plant of mangroves, Avicennia marinaena Forsk.Vierh.is widely distributed along the coast of South China Sea in China. It is a good material to study the molecular mechanism of heavy metal pollutant tolerance in mangrove plants. Based on the AmNramp3 fragment of heavy metal tolerance related gene obtained by suppression subtractive hybridization, a gene specific primer was designed. The length of cDNA was 1 958 bp.The expression of AmNramp3 in leaves, stems and roots of white bone soil under cadmium stress was detected by real-time fluorescence quantitative PCR qRT-PCR.The expression of AmNramp3 in leaves, stems and roots was higher than that in leaves and stems. But there was a rapid response in stem and root. The open reading frame of the gene was predicted and analyzed by bioinformatics, which was located at 1 527 BP, encoding 508 amino acids, and the structure of AmNramp3 protein was predicted and analyzed. It was found that its amino acid sequence did not contain a signal peptide, indicating that it was not a secretory protein, but it had a very complex transmembrane structure and was a transmembrane protein. It may be closely related to the transmembrane transport of cadmium, which lays the foundation for further analysis of the molecular mechanism of the function of the gene.
【作者单位】：厦门大学环境与生态学院;四川农业大学生命科学学院;
【基金】：国家自然科学基金(31530008,31370516)
【分类号】：Q943.2

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