| 野生二粒小麦低分子量谷蛋白基因序列分析 |
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| 引用本文： | 伍碧华,路洁霏,胡喜贵,李平,颜泽洪,刘登才,郑有良.野生二粒小麦低分子量谷蛋白基因序列分析[J].四川农业大学学报,2008,26(4). |
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| 作者姓名： | 伍碧华 路洁霏 胡喜贵 李平 颜泽洪 刘登才 郑有良 |
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| 作者单位： | 四川农业大学,小麦研究所,四川,温江,611130;四川农业大学,教育部作物基因资源与遗传改良重点实验室,四川,雅安,625014 |
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| 基金项目： | 国家自然科学基金，四川省教育厅重点项目，四川省学术与技术带头人培养基金 |
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| 摘 要： | 根据低分子量谷蛋白亚基(LMW-GS)基因编码区保守序列设计引物,用PCR方法从蛋白质含量低至13.17%的D81和高达27.20%的D42 2份野生二粒小麦(Triticumdicoccoides)中克隆得到2个LMW-GS基因序列LMW-D81和LMW-D42(GenBank上的序列号分别为FJ461691和FJ461690)。它们具有小麦低分子量谷蛋白基因的典型结构特征,其长度分别为1053bp和1011bp,并分别编码350和336个氨基酸残基的成熟蛋白。LMW-D42和LMW-D81的氨基酸序列估算分子量分别为38kDa和39kDa,说明二者均为C型亚基编码基因。LMW-D42和LMW-D81的N-末端序列都为METSHIP-,表明这2个C型亚基编码基因归属LMW-m型。同源性比对和聚类分析揭示,LMW-D81和LMW-D42均属于Glu-B3位点编码基因。LMW-D42和LMW-D81的核苷酸序列和推导的氨基酸序列一致性分别为93.94%和92.57%。与LMW-D81相比,LMW-D42除发生了22处间断性的碱基替换外,还存在一段42个碱基的缺失。对推导氨基酸序列进行的二级结构预测显示,LMW-D81和LMW-D42的蛋白质二级结构高度一致。其α-螺旋主要位于信号肽和C-末端,少量的α-螺旋和不规则卷曲构成了N-末端。大多数不规则卷曲位于重复区,仅有的一段β-折叠则出现在C-末端。同时,它们的编码区均具有分布一致的8个半胱氨酸残基,且第一和第七个半胱氨酸残基均位于无规则卷曲中。这些结构特点对小麦加工品质改良具有一定意义。
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| 关 键 词： | 野生二粒小麦 低分子量谷蛋白 基因克隆 序列分析 二级结构 |
Isolation and Characterization of Two Low Molecular Weight Glutenin Subunit Genes from Triticum dicoccoides |
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| WU Bi-hua,LU Jie-fei,HU Xi-gui,LI Ping,YAN Ze-hong,LIU Deng-cai,ZHENG You-liang.Isolation and Characterization of Two Low Molecular Weight Glutenin Subunit Genes from Triticum dicoccoides[J].Journal of Sichuan Agricultural University,2008,26(4). |
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| Authors: | WU Bi-hua LU Jie-fei HU Xi-gui LI Ping YAN Ze-hong LIU Deng-cai ZHENG You-liang |
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| Abstract: | In this paper,PCR for the genomic DNA from Triticum dicoccoides was conducted by using a pair of primers designed according to the conservative domain from the known low molecular weight glutenin subunit genes.The two LMW-GS genes,LMW-D81 and LMW-D42(GenBank accession number FJ461691 and FJ461690),were obtained from the accession D81 with the low protein content at 13.17% and D42 with the high protein content at 27.20%,respectively.LMW-D42 and LMW-D81 possessed the nucleotide sequences with 1011 bp and 1053 bp,and the deduced amino acid sequences with 336 and 350 amino acid residues,respectively.LMW-D42 and LMW-D81 belonged to C type genes owing to their molecular weight about 38 kDa and 39 kDa,respectively,as well as to LMW-m type genes based on the predicted protein sequences of N-terminal domain with METSHIP-.Meanwhile,LMW-D42 and LMW-D81 being allelic Glu-B3 genes were implied by homologous and cluster analyses.The similarities between their sequences were 93.94% in nucleotide and 92.57% in deduced amino acid,respectively.A single deletion for 42 bp occurred in LMW-D42 besides the substitutes at 22 interval sites,compared with LMW-D81.The predicted secondary structures of the amino acid sequences deduced from the two genes were similar.The signal peptide and C-terminal region consisted of many α-helixs.The N-terminal region was formed by a short section of both α-helixs and coils.The repetitive region was mainly occupied by coil.A single β-sheet structure was observed only in C-terminal region.Their amino acid sequences had 8 cysteine residues with similar distribution,and the first and seventh cysteine residues located in the coils.So,these structural characters of LMW-D42 and LMW-D81 might be of importance to quality improvement of bread wheat. |
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| Keywords: | Triticum dicoccoides low molecular weight glutenin gene cloning sequence analysis secondary structure |
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