欢迎访问作物学报，今天是
作物学报

作物学报 ›› 2010, Vol. 36 ›› Issue (3): 526-532.doi: 10.3724/SP.J.1006.2010.00526

• 研究简报 • 上一篇    下一篇

陕253γ-醇溶蛋白基因的克隆与序列分析

王明霞1，高翔1,2,*，陈其皎1,2，董剑1,2，赵万春1,2，李艳亮1，李敏1，陈瑞佶1，庞红喜1,2，李哲清1,2   

  1. 1西北农林科技大学农学院，陕西杨凌712100；2陕西省小麦工程技术研究中心/陕西省小麦新品种培育工程研究中心，陕西杨凌712100
  • 收稿日期:2009-09-24 修回日期:2009-12-08 出版日期:2010-03-12 网络出版日期:2010-01-22
  • 通讯作者: GAO Xiang,E-mail: gx@nwsuaf.edu.cn, Tel: 029-87082630
  • 基金资助:

    本研究由陕西省‘13115’科技创新工程重大项目（2007ZDKG-01）和现代农业产业技术体系建设专项（NYCYTX-001）资金资助。

Gene Cloning and Sequence Analysis of γ-Gliadin Genes from Wheat Cultivar Shann 253

WANG Ming-Xia1,GAO Xiang1,2,*,CHEN Qi-Jiao1,2,DONG Jian1,2,ZHAO Wan-Chun1,2, LI Yan-Liang1,LI Min1,CHEN Rui-Ji1,PANG Hong-Xi1,2,LI zhe-Qing1,2,LIU Jun1,2   

  1. 1 College of Agronomy, Northwest A&F University, Yangling 712100; 2 Wheat Engineering Research Center of Shaanxi Province / New varieties cultivation of wheat Engineering Research Center of Shaanxi Province; Yangling 712100, China
  • Received:2009-09-24 Revised:2009-12-08 Published:2010-03-12 Published online:2010-01-22
  • Contact: 高翔, E-mail: gx@nwsuaf.edu.cn, Tel: 029-87082630

PDF

1961

被引次数

2

摘要：

针对γ-醇溶蛋白基因家族成员，设计了覆盖其启动子及全长编码区的3对特异引物，从强筋小麦品种陕253中克隆了8条1 000 bp左右的片段(GenBank登录号为GQ871770~GQ871777)。该片段群包含典型醇溶蛋白亚基的完整编码序列，并在重复区存在丰富的插入/缺失(InDel)；推导的氨基酸序列显示，8个基因均具有γ-醇溶蛋白亚基的典型结构特征，其中GQ871771为假基因，4条序列(GQ871770、GQ871772、GQ871776和GQ87177)具有9个半胱氨酸残基；启动子区序列分析表明，GQ871770、GQ871772、GQ871774和GQ871776在胚乳框存在6处SNP变异，其中两处变异发生于GCN4基序内，利用WebLogo3在线构建了储藏蛋白更具代表意义的30 bp保守胚乳盒模式。进化分析证实克隆序列属于γ-醇溶蛋白基因家族成员。

关键词: 普通小麦, γ-醇溶蛋白, 基因克隆, 序列分析

Abstract:

Eight γ-gliadin genes (GenBank accession number GQ87170 to GQ87177) were cloned from wheat (Triticum aestivum L.) cultivar Shaan 253 using three sets of specific primers, which were designed according to the known γ-gliadin gene family. All of the eight genes possess the typical structure of γ-gliadin subunits and many sequences of insent/indel in the repeat region. The deduced amino acid sequences showed that GQ871771 is a pseudogene, and nine cysteine residues existed in GQ871770, GQ871772, GQ871776, and GQ87177. Promoter sequences analysis indicated that there were six SNPs in endosperm box of GQ871770, GQ871772, GQ87174, and GQ871776, and two of them appeared in the GCN4-like motif. A model for endosperm box of storage protein with better representation was constructed using the online tool WebLogo3. Phylogenetic analysis also confirmed that the cloned sequences belong to γ-gliadin gene family.

Key words: Common wheat, γ-gliadin gene, Gene clone, Sequence analysis

[1]        Bushuk W. Wheat cultivar identification by gliadin electrophoregrams. Can J Plant Sci, 1978, 58: 505−515

[2]        Harbred N P, Bartels D, Thompson R D. Analysis of the gliadin multigene loci in bread wheat using nulisomic tetrasomic lines. Mol Gen Genet, 1985, 198: 234−242

[3]        Wrifley C W, Robinson P J, Williams W T. Association between individual gliadin proteins and quality, agronomic and morphological attributes of wheat cultivars. Aust Agric Res, 1982, 33: 409−418

[4]        Redaelli R. Two-dimensional mapping of gliadins using biotypes and null mutants of common wheat cultivar Saratovskaya 29. Hereditas, 1994, 121: 131−137

[5]        Peter R, Nigel G. Cereal seed storage proteins: structures properties and role in grain utilization. J Exp Pathol, 2002, 53: 370

[6]        Yan Y-M(晏月明), Ru Y-Y(茹岩岩), Yu J-Z(余建中), Liu G-T(刘广田), Prodanovic S. Analysis of gliadin allele composition at Gli-1 and Gli-2 loci in Chinese wheat cultivars. J Agric Biotechnol (农业生物技术学报), 2000, 6(1): 23−27 (in Chinese with English abstract)

[7]        Branlard G, Dardevet M. Diversity of grain proteins and bread wheat quality I correlation between gliadin bands and flour quality characteristics. Cereal Sci, 1985,13: 329−343

[8]        Payne P I. Genetics of wheat storage proteins and the effect of allelic variation on bread-making quality. Annu Rev Genet, 1987, 38: 141−153

[9]        Xu Z-F (徐兆飞), Zhang H-Y (张惠叶), Zhang D-Y (张定一). Wheat Quality and Its Improvement (小麦品质及其改良). Beijing: Meteorological Press, 2000. pp 47−51 (in Chinese)

[10]     Murray H G, Thompson W F. Rapid isolation of high molecular weight DNA. Nucl Acids Res, 1980, 8: 4321−4325

[11]     Tamura K, Dudley J, Nei M, Kumar S. MEGA4: Molecular Evolutionary Genetics Analysis (MEGA) software version 4.0. Mol Biol Evol, 2007, 24: 1596−1599

[12]     Higo K, Ugawa Y, Iwamoto M, Korenaga T. Plant cis-acting regulatory DNA elements (PLACE) database:1999. Nucl Acids Res, 1999, 27: 297−300

[13]     Crooks G E, Hon G, Chandonia J M, Brenner S E. WebLogo: A sequence logo generator. Genome Res, 2004, 14:1188−1190

[14]     Schneider T D, Stephens R M. Sequence Logos: A new way to display consensus sequences. Nucl Acids Res, 1990, 18: 6097−6100

[15]     Bartels D, Thompson R D. The characterization of cDNA clones coding for wheat storage proteins. Nucl Acids Res, 1983, 11: 2961−2977

[16]     Kasarda D D, Okita T W, Bernardin J E. Nucleic acid and amino acid sequences of α-type gliadins from wheat. Proc Natl Acad Sci, 1984, 81: 4712-4716

[17]     Jackson E A, Morel M H, T Sontag-Strohm. Proposal for combination the classification system of alleles of Gli-1 and Gli-3 loci in bread wheat. Genet Breed, 1996, 50: 321-336

[18]     Anderson O D, Hisa C C, Torres V. The wheat γ-gliadin genes: characterization of ten new sequences and further understanding of γ-gliadin gene family structure. Theor Appl Genet, 2001, 103: 323−330

[19]     Yan Y-M(晏月明), Liu G-T(刘广田), Prodanovic S, Zoric D. Genetic and quality improvement on wheat gliadin. Tritical Crops(麦类作物), 1998, 18(1): 1−4 (in Chinese with English abstract)

[20]     Sun C-X(孙崇荣), Huang W-D(黄伟达). Expression regulation of storage genes in seeds of wheat. Commun Plant Physiol (植物生理学通讯), 1995, 31(1): 71−73 (in Chinese)

[21]     Thomas M S, Flavell R B. Identification of an enhancer element for the endosperm specific expression of high molecular weight glutenin. Plant Cell, 1990, 2: 1171−1180

[22]     Vellanoveth R L, Okita T W. Analysis of nuclear proteins interacting with a wheat gliadin seed storage protein gene. Plant Mol Biol, 1993, 22: 25−41

[23]     Gupta R B, MacRcitchie F. Allelic variation at glutenin subunit and gliadin loci, Glu-1, Glu-3, Gli-1 of common wheat: II. Biochemical basis of the allelic effects on dough properties. J Cereal Sic, 1994, 19: 19−29

[24]     Vensel W H, Tarr G E, Kasarda D D. C-terminal and internal sequence of a low molecular weight (LMW-s) type of glutenin subunit. Cereal Chem, 1995, 72: 356−359
Shewry P R, Tatham A S. The proclaim storage proteins of cereal seeds: structure and evolution. Biochem J, 1990, 267: 1−12
[1] 左同鸿, 张贺翠, 曾静, 朱利泉. 甘蓝自交不亲和相关基因BoPUB3L的克隆与表达分析[J]. 作物学报, 2026, 52(6): 1698-1710.
[2] 余开航, 周洪斌, 罗亮扎, 王玫郦, 姜瑞梅, 董陈文华, 李仕金, 毛孝强, 陈升位. 大麦亮氨酸富集重复型类受体激酶基因HvLRR-RLK-510的克隆和表达分析[J]. 作物学报, 2026, 52(2): 421-432.
[3] 薛晓菲, 戴云静, 李熙林, 丁艳艳, 王翔, 雷长英, 韩焕勇, 贺道华. 陆地棉杜松烯合酶基因GhCDN10的特征及其在棉酚合成中功能分析[J]. 作物学报, 2025, 51(8): 2060-2076.
[4] 王林, 陈晓雨, 张文梦龙, 汪思琦, 程冰云, 程靖秋, 潘锐, 张文英. 大麦HvMYB2分子特性及响应干旱胁迫的功能分析[J]. 作物学报, 2025, 51(4): 873-887.
[5] 张正康, 苏延红, 阮孙美, 张敏, 张攀, 张慧, 曾千春, 罗琼. 疣粒野生稻中OgXa13的克隆和功能研究[J]. 作物学报, 2025, 51(2): 334-346.
[6] 王玉娇, 王永乐, 添长久, 郁春旺, 吕佳斌, 朱加保. 薏苡VQ4基因的克隆及耐盐性初步分析[J]. 作物学报, 2025, 51(12): 3198-3210.
[7] 胡瑶洁, 刘亚萍, 郑君妍, 韩婷, 马伯军, 叶亚峰, 刘斌美, 陈析丰. 水稻类病变早衰突变体lmes6的表型鉴定与基因定位[J]. 作物学报, 2024, 50(11): 2764-2774.
[8] 刘佳, 龚方仪, 刘亚西, 颜泽洪, 钟晓英, 陈厚霖, 黄林, 伍碧华. 野生二粒小麦主要农艺特性融入普通小麦的全基因组关联分析[J]. 作物学报, 2023, 49(5): 1184-1196.
[9] 靖小菁, 杨新笋, 靳晓杰, 刘意, 雷剑, 王连军, 柴沙沙, 张文英, 焦春海. 甘薯蔓割病(Fusarium oxysporum f. sp. batatas)相关基因IbMAPKK9的克隆与特性分析[J]. 作物学报, 2023, 49(12): 3289-3301.
[10] 王穆穆, 何艳芳, 郑永胜, 王晖, 王丽媛, 王东建, 张晗, 李汝玉. 水稻落粒基因SH8的精细定位与克隆[J]. 作物学报, 2022, 48(8): 1948-1956.
[11] 崔连花, 詹为民, 杨陆浩, 王少瓷, 马文奇, 姜良良, 张艳培, 杨建平, 杨青华. 2个玉米ZmCOP1基因的克隆及其转录丰度对不同光质处理的响应[J]. 作物学报, 2022, 48(6): 1312-1324.
[12] 周慧文, 丘立杭, 黄杏, 李强, 陈荣发, 范业赓, 罗含敏, 闫海锋, 翁梦苓, 周忠凤, 吴建明. 甘蔗赤霉素氧化酶基因ScGA20ox1的克隆及功能分析[J]. 作物学报, 2022, 48(4): 1017-1026.
[13] 杨陆浩, 王立建, 孙广华, 王少瓷, 崔连花, 陈昌, 宋梅芳, 张艳培, 姜良良, 杨建平, 王晨阳. 栽培黑麦光敏色素PHYA、PHYB和PHYC基因转录丰度对不同光质处理的响应[J]. 作物学报, 2022, 48(12): 3057-3070.
[14] 谢琴琴, 左同鸿, 胡燈科, 刘倩莹, 张以忠, 张贺翠, 曾文艺, 袁崇墨, 朱利泉. 甘蓝自交不亲和相关基因BoPUB9的克隆及表达分析[J]. 作物学报, 2022, 48(1): 108-120.
[15] 唐锐敏, 贾小云, 朱文娇, 印敬明, 杨清. 马铃薯热激转录因子HsfA3基因的克隆及其耐热性功能分析[J]. 作物学报, 2021, 47(4): 672-683.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备15008789号-2