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花生金属硫蛋白基因AhMT3a的克隆及其表达
引用本文:全先庆,高成香,王兴军,赵传志.花生金属硫蛋白基因AhMT3a的克隆及其表达[J].生物技术通报,2012,0(3):75-79.
作者姓名:全先庆  高成香  王兴军  赵传志
作者单位:1. 临沂大学生命科学学院,临沂,276005
2. 潍坊护理职业学院,青州,262500
3. 山东省农业科学院高新技术研究中心,济南,250100
基金项目:山东省自然科学基金资助项目
摘    要:以鲁花14号花生为材料,从花生cDNA文库和基因组中筛选和克隆了花生的金属硫蛋白基因AhMT3a。该基因全长785 bp,有2个内含子,开放阅读框由201个碱基组成,编码66个氨基酸,其中包含13个半胱氨酸(Cys),预测其分子量为6.83kD,等电点为4.59。运用生物信息学手段对AhMT3a蛋白的信号肽、跨膜区、亚细胞定位和疏水性进行了预测。与拟南芥、棉花和草莓等植物type 3 MTs的序列比对结果表明,花生和其他不同植物的MT3在氨基酸序列上具有较高的同源性,从系统发育树中可以看出AhMT3a和蒿麦的金属硫蛋白亲缘关系较近。半定量RT-PCR和芯片杂交结果显示花生AhMT3a在花中表达量最高,在种子中表达量最低;在ABA、NaCl及PEG等不同处理下,表达量变化不大。

关 键 词:花生(Arachis  hypogaea  L.)  金属硫蛋白  基因克隆  表达分析

Cloning and Expression Analysis of AhMT3a in Peanut (Arachis hypogaea L.)
Quan Xianqing , Gao Chengxiang , Wang Xingjun , Zhao Chuanzhi.Cloning and Expression Analysis of AhMT3a in Peanut (Arachis hypogaea L.)[J].Biotechnology Bulletin,2012,0(3):75-79.
Authors:Quan Xianqing  Gao Chengxiang  Wang Xingjun  Zhao Chuanzhi
Institution:1College of Life Science,Linyi University,Linyi 276005;2High-Tech Research Center,Shandong Academy of Agricultural Sciences, Jinan 250100;3Weifang College of Nursing Occupation,Qingzhou 262500)
Abstract:A type 3 MT gene AhMT3a was screened from cDNA library of peanut,and the full-length genomic DNA was cloned by PCR.The length of AhMT3a is 785 bp with two introns.The ORF is 201 bp and encodes 66 amino acids which contains 13 cysteines.The predicted molecular weight of AhMT3a is 683 kD.The signal peptide,transmembrane area,sub-cellular location and hydrophobicity were also predicted by bioinformatic software.The phylogenetic tree showed that AhMT3a was closely related to cotton MT3 protein.Semi-quantitative RT-PCR and microarray results showed that AhMT3a expressed in roots,stems,flowers and seeds while the expression level in flowers was high.The expression level of AhMT3a did not change significantly upon ABA,NaCl and PEG treatment.
Keywords:Peanut(Arachis hypogaea L  ) Metallothionein Gene cloning Expression analysis
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