花生膜联蛋白基因AhAnn1的克隆与表达分析

发布时间：2018-03-15 08:50

本文选题：花生　切入点：膜联蛋白基因　出处：《华北农学报》2017年03期 　论文类型：期刊论文

【摘要】：为挖掘花生抗逆境胁迫相关基因,克隆抗逆相关膜联蛋白Annexin基因,以花生品种花育25号为试验材料,从已知抑制消减杂交文库中获得花生膜联蛋白Annexin类似基因片段,通过RACE技术获得Annexin基因5'-RACE片段。对5'-RACE序列和抑制消减杂交文库中已知基因片段进行拼接,设计特异引物通过逆转录-聚合酶链式反应(RT-PCR)扩增得到该基因,命名为AhAnn1。结果表明,AhAnn1全长为1 277 bp,开放阅读框为951 bp。根据编码区预测AhAnn1编码一条316个氨基酸组成的多肽,预测分子量为36.10 k Da,等电点为7.07。预测该基因编码的蛋白含有膜联蛋白Annexin保守结构域,定位于细胞质中。蛋白序列多重比对和系统发育分析表明,花生与大豆、苜蓿、鹰嘴豆等豆科植物中的膜联蛋白相似性最高,亲缘关系最近。荧光实时定量PCR结果显示,AhAnn1在根系和叶片中的表达量随干旱胁迫程度的增加而增加。由此推测AhAnn1是一种干旱胁迫应答基因,在花生逆境调控中发挥作用。结果为进一步研究花生AhAnn1基因的功能和花生抗逆境胁迫相关分子机理提供了理论基础。
[Abstract]:For mining peanut stress resistance related genes, cloning resistance related annexin Annexin gene, peanut cultivar Huayu 25 as test material, known from the suppression subtractive hybridization library in peanut annexin Annexin like gene fragment by RACE from Annexin gene 5'-RACE fragment. The 5'-RACE sequence and known suppression subtractive hybrid library gene fragments were spliced with specific primers by reverse transcriptase polymerase chain reaction (RT-PCR) amplified the gene, named AhAnn1.. The results showed that the full-length AhAnn1 was 1277 BP, the open reading frame of 951 bp. encoding a AhAnn1 polypeptide according to 316 amino acids predicted encoding region, a predicted molecular weight of 36.10 K Da, the isoelectric point of 7.07. to predict the gene encoding a protein containing annexin Annexin conserved domain, located in the cytoplasm. The protein multiple sequence alignment and system Fertility analysis showed that peanut and soybean, alfalfa, chickpeas and other legumes annexin the highest similarity, the closest genetic relationship. Real time fluorescent quantitative PCR results showed that the expression of AhAnn1 in roots and leaf weight with the increase of drought stress increased. It suggested that AhAnn1 is a kind of drought stress responsive genes. Play a role in the regulation of stress in peanut. The results provide a theoretical basis for further research on the molecular mechanism of stress related peanut AhAnn1 gene function and peanut anti adversity.

【作者单位】：山东省花生研究所;
【基金】：山东省农业科学院青年英才培养计划项目山东省优秀中青年科学家科研奖励基金项目(BS2012NY010) 国家自然科学基金项目(31201171) 山东省现代农业产业技术体系花生创新团队(SDAIT-04-06)
【分类号】：Q943.2;S565.2

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