PDF下载 +分享

微信公众号：大豆科学

[1]张承琪,王怡,夏成林,等.大豆F-box基因Glyma08g11030启动子克隆及植物表达载体构建[J].大豆科学,2018,37(02):179-184.[doi:10.11861/j.issn.1000-9841.2018.02.0179]
　ZHANG Cheng-qi,WANG Yi,XIA Cheng-lin,et al.Isolation of Glyma08g11030 Promoter from F-box Gene of Glycine max and Construction of Its Plant Expression Vectors[J].Soybean Science,2018,37(02):179-184.[doi:10.11861/j.issn.1000-9841.2018.02.0179]

点击复制

大豆F-box基因Glyma08g11030启动子克隆及植物表达载体构建

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第37卷期数: 2018年02期页码: 179-184 栏目: 出版日期: 2018-03-20

Title:: Isolation of Glyma08g11030 Promoter from F-box Gene of Glycine max and Construction of Its Plant Expression Vectors

作者:: 张承琪; 王怡; 夏成林; 于月华; 倪志勇; （新疆农业大学农学院,新疆乌鲁木齐 830052）

Author(s):: ZHANG Cheng-qi; WANG Yi; XIA Cheng-lin; YU Yue-hua; NI Zhi-yong; （College of agronomy, Xinjiang Agricultural University, Urumqi 830052, China）

关键词:: 大豆; 启动子; 克隆; 植物表达载体

Keywords:: Soybean; Promoter; Clone; Plant expression vector

分类号:: S565.1；Q78

DOI:: 10.11861/j.issn.1000-9841.2018.02.0179

文献标志码:: A

摘要:: 本研究以大豆Williams 82的基因组DNA为原始材料，根据前期预测的Glyma08g11030启动子序列用DNAMAN8.0设计引物，通过常规PCR方法克隆获得了Glyma08g11030基因上游长度为3 000 bp的启动子序列。在PlantCARE植物启动子区域在线预测网站上查询，预测的结果显示Glyma08g11030启动子序列里含有数量众多、功能各异的顺式作用元件。其中以TATA-box和CAAT-box这两个基本的顺式作用元件最多，此外还有许多与应答相关的顺式作用元件，如干旱、热、光、激素等。根据这些顺式作用元件在Glyma08g11030启动子序列中的位置对启动子序列进行截短并分别设计引物，采用PCR方法成功截出了3段长度分别为2 492，1 185和342 bp的启动子序列,将3段序列替换pCAMBIA3301载体双酶切出的CaMV35S启动子序列，并构建了3个启动子序列驱动GUS植物融合表达载体，从而得到含不同顺式作用元件的启动子序列。研究结果将为以后分析Glyma08g11030启动子的功能和作用机理提供前期准备。

Abstract:: Based on the predicted sequence of Glyma08g11030 promoter region, primers were designed based on promoter sequence. We cloned 3 000 bp fragments from soybean Williams 82 genome DNA by PCR method.The promoter sequence was placed on the PlantCARE plant promoter region to predict online website queries. The predicted results showed that Glyma08g11030 promoter sequence contained a large number of cis-acting elements with various functions. The two basic cis-acting elements of TATA-box and CAAT-box were the most, and there were many cis-acting elements responding to it, such as drought, heat, light and hormones. According to the position of these cis-acting elements in the Glyma08g11030 promoter sequence, the primer sequences were truncated and the primers were designed respectively. Three truncated promoters were cloned successfully by PCR method and their length were 2 492, 1 185 and 342 bp. We constructed three GUS plant fusion expression vectors by replacing CaMV35S promoter of pCAMBIA3301 and obtained the promoter sequences with different cis-acting elements. These results would be helpful for the further research of functional characterization of cis-acting elements and regulation mechanism in Glyma08g11030 promoter.

参考文献/References:

［1］宋阳, 王丕武, 张学明, 等. 大豆豆荚特异性启动子的克隆及功能分析［J］.西北农林科技大学学报, 2014, 42(10): 63-80. (Song Y, Wang P W, Zhang X M, et al. Cloning and functional analysis of soybean pod specific promoter［J］. Journal of Northwest A & F University, 2014, 42(10): 63-80.)?

［2］Zhu J K. Salt and drought stress signal transduction in plants［J］. Annual Review of Plant Biology, 2002, 53(53): 247.?

［3］Kizis D, Pagès M. Maize DRE-binding proteins DBF1 and DBF2 are involved in rab17 regulation through the drought-responsive element in an ABA-dependent pathway［J］. Plant Journal for Cell & Molecular Biology, 2002, 30(6): 679-689.?

［4］翟莹, 张军, 赵艳, 等. 大豆转录因子GmERF5启动子的克隆及活性分析［J］. 生物技术通报, 2014(1): 100-104. (Zhai Y, Zhang J, Zhao Y, et al. Cloning and activity analysis of soybean GmERF5 promoter［J］. Biotechnology bulletin, 2014(1): 100-104.)

［5］Xiao W, Jang J. F-box proteins in Arabidopsis ［J］. Trends in Plant Science, 2000, 5(11): 454.

［6］Zhao Z, Zhang G, Zhou S, et al. The improvement of salt tolerance in transgenic tobacco by overexpression of wheat F-Box gene TaFBA1［J］. Plant Science, 2017, 259: 71-85.?

［7］Zhou S M, Kong X Z, Kang H H, et al. The involvement of wheat F-Box protein gene TaFBA1 in the oxidative stress tolerance of plants［J］. Plos One, 2015, 10(4): e0122117.?

［8］Kong X, Zhou S, Yin S, et al. Stress-inducible expression of an F-box Gene TaFBA1 from wheat enhanced the drought tolerance in transgenic tobacco plants without impacting growth and development［J］. Frontiers in Plant Science, 2016, 7(135): 01295.?

［9］Li Y, Jia F, Yu Y, et al. The SCF E3 ligase AtPP2-B11 plays a negative role in response to drought stress in Arabidopsis［J］. Plant Molecular Biology Reporter, 2014, 32(5): 943-956.?

［10］Jia F, Wang C, Huang J, et al. SCF E3 ligase PP2-B11 plays a positive role in response to salt stress in Arabidopsis［J］. Journal of Experimental Botany, 2015, 66(15): 4683.?

［11］罗中钦. 大豆逆境胁迫相关microRNA的发掘与验证［D］. 北京: 中国农业科学院, 2012. (Luo Z Q. Exploration and verification of stress-responsive microRNA from soybean［D］. Beijing: Chinese Academy of Agricultural Sciences, 2012.)

［12］Zhu L P, Yu Z, Zou C X, et al. Plant stress-inducible promoters and their function［J］. Hereditas, 2010, 32(3): 229.?

［13］倪志勇. 大豆抗逆相关miR169c及其靶位点GmNFYA3和miR394a的功能研究［D］. 北京: 中国农业科学院, 2013. (Ni Z Y. Functional Study of Stress-Responsive miR169c and its target gene GmNFYA3 and miR394a from Soybean ［D］. Beijing: Chinese Academy of Agricultural Sciences, 2013.)

［14］Postel D, Vanlemmens P, Gode P, et al. PlantCARE, a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences［J］. Nucleic Acids Research, 2002, 30(1): 325-327.?

［15］李冬梅, 王涛, 陈薇, 等. 大豆Glyma03g24460基因功能预测及表达分析［J］. 基因组学与应用生物学, 2016(3): 687-691. (Li D M, Wang T, Chen W, et al. Functional prediction and expression analysis of Glyma03g24460 gene from soybean［J］. Genomics and Applied Biology, 2016(3): 687-691)

［16］倪志勇, 于月华, 任燕萍, 等. 大豆gma-miR169c启动子的克隆及植物表达载体构建［J］. 分子植物育种, 2015, 13(2): 287-293. (Ni Z Y, Yu Y H, Ren Y P, et al. Isolation of gma-mi R169c promoter form Glycine max and construction of its plant expression vectors［J］. Molecular Plant Breeding, 2015, 13(2): 287-293.)

［17］孙晓丽, 李勇, 才华, 等. 拟南芥bZIP1转录因子通过与ABRE元件结合调节ABA信号传导［J］. 作物学报, 2011, 37(4): 612-619. (Sun X L, Li Y, Cai H, et al. Arabidopsis bZIP1 transcription factor binding to the ABRE cis-element regulates abscisic acid signal transduction［J］. Acta Agronomica Sinica, 2011, 37(4):612-619.)

［18］丑晓红, 王春燕. 玉米Zma-miR169i启动子的克隆及分析［J］. 分子植物育种, 2017(1): 17-22. (Chou X H, Wang C Y. Cloning and analysis of Zma-miR169i promoter in Zea mays［J］. Molecular Plant Breeding, 2017(1):17-22.)

［19］Kasuga M, Liu Q, Miura S. Improving plant drought, salt, and freezing tolerance by gene transfer of a single stress-inducible transcription factor1［J］. Nature Biotechnology, 1999, 17(3): 287-291.

相似文献/References:

[1]刘章雄,李卫东,孙石,等.1983～2010年北京大豆育成品种的亲本地理来源及其遗传贡献[J].大豆科学,2013,32(01):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
　LIU Zhang-xiong,LI Wei-dong,SUN Shi,et al.Geographical Sources of Germplasm and Their Nuclear Contribution to Soybean Cultivars Released during 1983 to 2010 in Beijing[J].Soybean Science,2013,32(02):1.[doi:10.3969/j.issn.1000-9841.2013.01.002]
[2]李彩云,余永亮,杨红旗,等.大豆脂质转运蛋白基因GmLTP3的特征分析[J].大豆科学,2013,32(01):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
　LI Cai-yun,YU Yong-liang,YANG Hong-qi,et al.Characteristics of a Lipid-transfer Protein Gene GmLTP3 in Glycine max[J].Soybean Science,2013,32(02):8.[doi:10.3969/j.issn.1000-9841.2013.01.003]
[3]王明霞,崔晓霞,薛晨晨,等.大豆耐盐基因GmHAL3a的克隆及RNAi载体的构建[J].大豆科学,2013,32(01):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
　WANG Ming-xia,CUI Xiao-xia,XUE Chen-chen,et al.Cloning of Halotolerance 3 Gene and Construction of Its RNAi Vector in Soybean (Glycine max)[J].Soybean Science,2013,32(02):12.[doi:10.3969/j.issn.1000-9841.2013.01.004]
[4]张春宝,李玉秋,彭宝,等.线粒体ISSR与SCAR标记鉴定大豆细胞质雄性不育系与保持系[J].大豆科学,2013,32(01):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
　ZHANG Chun-bao,LI Yu-qiu,PENG Bao,et al.Identification of Soybean Cytoplasmic Male Sterile Line and Maintainer Line with Mitochondrial ISSR and SCAR Markers[J].Soybean Science,2013,32(02):19.[doi:10.3969/j.issn.1000-9841.2013.01.005]
[5]卢清瑶,赵琳,李冬梅,等.RAV基因对拟南芥和大豆不定芽再生的影响[J].大豆科学,2013,32(01):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
　LU Qing-yao,ZHAO Lin,LI Dong-mei,et al.Effects of RAV gene on Shoot Regeneration of Arabidopsis and Soybean[J].Soybean Science,2013,32(02):23.[doi:10.3969/j.issn.1000-9841.2013.01.006]
[6]杜景红,刘丽君.大豆fad3c基因沉默载体的构建[J].大豆科学,2013,32(01):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
　DU Jing-hong,LIU Li-jun.Construction of fad3c Gene Silencing Vector in Soybean[J].Soybean Science,2013,32(02):28.[doi:10.3969/j.issn.1000-9841.2013.01.007]
[7]张力伟,樊颖伦,牛腾飞,等.大豆“冀黄13”突变体筛选及突变体库的建立[J].大豆科学,2013,32(01):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
　ZHANG Li-wei,FAN Ying-lun,NIU Teng-fei?,et al.Screening of Mutants and Construction of Mutant Population for Soybean Cultivar "Jihuang13”[J].Soybean Science,2013,32(02):33.[doi:10.3969/j.issn.1000-9841.2013.01.008]
[8]盖江南,张彬彬,吴瑶,等.大豆不定胚悬浮培养基因型筛选及基因枪遗传转化的研究[J].大豆科学,2013,32(01):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
　GAI Jiang-nan,ZHANG Bin-bin,WU Yao,et al.Screening of Soybean Genotypes Suitable for Suspension Culture with Adventitious Embryos and Genetic Transformation by Particle Bombardment[J].Soybean Science,2013,32(02):38.[doi:10.3969/j.issn.1000-9841.2013.01.009]
[9]王鹏飞,刘丽君,唐晓飞,等.适于体细胞胚发生的大豆基因型筛选[J].大豆科学,2013,32(01):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
　WANG Peng-fei,LIU Li-jun,TANG Xiao-fei,et al.Screening of Soybean Genotypes Suitable for Somatic Embryogenesis[J].Soybean Science,2013,32(02):43.[doi:10.3969/j.issn.1000-9841.2013.01.010]
[10]刘德兴,年海,杨存义,等.耐酸铝大豆品种资源的筛选与鉴定[J].大豆科学,2013,32(01):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
　LIU De-xing,NIAN Hai,YANG Cun-yi,et al.Screening and Identifying Soybean Germplasm Tolerant to Acid Aluminum[J].Soybean Science,2013,32(02):46.[doi:10.3969/j.issn.1000-9841.2013.01.011]
[11]赵艳,赵丹,刘博,等.大豆P39-1基因及其启动子表达方式的初步研究[J].大豆科学,2013,32(03):425.[doi:10.11861/j.issn.1000-9841.2013.03.0425]
　ZHAO Yan,ZHAO Dan,LIU Bo,et al.Preliminary Studies on the Expression Pattern of Soybean P39-1 Gene and Promoter[J].Soybean Science,2013,32(02):425.[doi:10.11861/j.issn.1000-9841.2013.03.0425]
[12]赵艳,祁宏英,范震宇,等.大豆酰基载体蛋白硫酯酶基因及其启动子表达方式的初步分析[J].大豆科学,2012,31(05):714.[doi:10.3969/j.issn.1000-9841.2012.05.005]
　ZHAO Yan,QI Hong-ying,FAN Zhen-yu,et al.Preliminary Analysis of the Expression Pattern of Soybean acyl-ACP-Thioesterase Gene and Promoter[J].Soybean Science,2012,31(02):714.[doi:10.3969/j.issn.1000-9841.2012.05.005]
[13]王林,韩飞,李爱科,等.不同加工工艺对大豆转基因成分及调控元件的影响[J].大豆科学,2011,30(01):136.[doi:10.11861/j.issn.1000-9841.2011.01.0136]
　WANG Lin,HAN Fei,LI Ai-ke,et al.Effect of Different Processing on Transgenic Ingredients and Regulatory Elements of Soybean[J].Soybean Science,2011,30(02):136.[doi:10.11861/j.issn.1000-9841.2011.01.0136]
[14]李永光,艾佳,王涛,等.大豆gma-miR1508a靶基因预测及功能分析[J].大豆科学,2014,33(04):483.[doi:10.11861/j.issn.1000-9841.2014.04.0479]
　LI Yong-guang,AI Jia,WANG Tao,et al.The Target Genes Prediction and Analysis of gma-miR1508a [J].Soybean Science,2014,33(02):483.[doi:10.11861/j.issn.1000-9841.2014.04.0479]
[15]郭伟,汪潇淋,王慧,等.GmAGL15基因启动子的克隆及序列分析[J].大豆科学,2008,27(05):727.[doi:10.11861/j.issn.1000-9841.2008.05.0727]
　GUO Wei,WANG Xiao-lin,WANG Hui,et al.Cloning and Sequence Analysis of GmAGL15 Gene Promoter from Soybean[J].Soybean Science,2008,27(02):727.[doi:10.11861/j.issn.1000-9841.2008.05.0727]
[16]倪志勇,于月华,陈全家,等.大豆gma-miR1514b生物信息学分析及人工microRNA植物表达载体构建[J].大豆科学,2016,35(01):46.[doi:10.11861/j.issn.1000-9841.2016.01.0046]
　NI Zhi-yong,YU Yue-hua,CHEN Quan-jia,et al.Bioinformatic Analysis of gma-miR1514b and Construction of Its Artificial miRNA Plant Expression Vectors[J].Soybean Science,2016,35(02):46.[doi:10.11861/j.issn.1000-9841.2016.01.0046]
[17]倪志勇,于月华,陈全家,等.大豆gma-miR1510a生物信息学分析及人工microRNA植物表达载体构建[J].大豆科学,2016,35(02):239.[doi:10.11861/j.issn.1000-9841.2016.02.0239]
　NI Zhi-yong,YU Yue-hua,CHEN Quan-jia,et al.Bioinformatic Analysis of gma-miR1510a and Construction of Its Artificial MiRNA Plant Expression Vectors[J].Soybean Science,2016,35(02):239.[doi:10.11861/j.issn.1000-9841.2016.02.0239]
[18]金玲,张浩,王松明,等.大豆GmAMS基因及其启动子的克隆和表达分析[J].大豆科学,2019,38(06):889.[doi:10.11861/j.issn.1000-9841.2019.06.0889]
　JIN Ling,ZHANG Hao,WANG Song-ming,et al.Cloning and Expression Analysis of GmAMS Gene and Its Promoter in Soybean[J].Soybean Science,2019,38(02):889.[doi:10.11861/j.issn.1000-9841.2019.06.0889]
[19]张军,尹珺伊,何佳琦,等.大豆GmDof4和GmDof11基因的非生物胁迫诱导表达及启动子分析[J].大豆科学,2021,40(02):195.[doi:10.11861/j.issn.1000-9841.2021.02.0195]
　ZHANG Jun,YIN Jun-yi,HE Jia-qi,et al.Abiotic Stress Induced Expression and Promoter Prediction of GmDof4 and GmDof11 Gene in Soybean[J].Soybean Science,2021,40(02):195.[doi:10.11861/j.issn.1000-9841.2021.02.0195]
[20]胡慧敏,蔡婉菡,穆可彬,等.大豆ABA信号途径GmCDPK SK5基因异源表达探究[J].大豆科学,2021,40(06):737.[doi:10.11861/j.issn.1000-9841.2021.06.0737]
　HU Hui-min,CAI Wan-han,MU Ke-bin,et al.Study on the Heterologous Expression of GMCDPK SK5 Gene in Soybean ABA Signaling Pathway[J].Soybean Science,2021,40(02):737.[doi:10.11861/j.issn.1000-9841.2021.06.0737]

备注/Memo

收稿日期：2018-02-01

基金项目：国家自然科学基金（31660295）；中国博士后科学基金（2015M582741）；新疆维吾尔族自治区科研创新项目（XJGRI2017063）。

第一作者简介：张承琪（1992-），男，硕士，主要从事植物逆境分子生物学研究。E-mail：576760973@qq.com。

通讯作者：倪志勇（1981-），男，博士，副教授，主要从事植物逆境分子生物学研究。E-mail：nizhiyong@126.com。

更新日期/Last Update: 2018-04-02