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苎麻BnbZIP1转录因子基因的克隆与表达特征分析 被引量:7

Cloning and Characterization of the BnbZIP1 Transcription Factor Gene from Ramie (Boehmeria nivea L.)
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摘要 【目的】克隆苎麻转录因子基因BnbZIP1,分析其序列及表达特征,同时进行原核表达以及亚细胞定位分析。【方法】根据苎麻转录组测序中的Unigene48047片段序列,采用RT-PCR结合RACE技术克隆该基因的全长cDNA序列,并进行生物信息学分析;利用Real-time PCR分析该基因在不同组织和不同胁迫条件下表达特征;构建原核表达载体,进行原核表达;构建含EGFP的融合表达载体,观察其亚细胞定位情况。【结果】苎麻BnbZIP1的cDNA全长为2 071 bp,开放读码框为1 407 bp,编码468个氨基酸,推导该多肽的等电点和分子量为7.12和51.57kD,与毛果杨(XP_002307972)的bZIP基因氨基酸序列的相似性最高,达到93%;IPTG诱导产生的重组蛋白相对分子量约为52 kD,与理论值一致;亚细胞定位分析表明其定位在细胞核中;荧光定量PCR分析表明,该基因在苎麻根、茎、茎尖、叶片、雌花和雄花中均有表达,其中,在雄花中表达最高,在根中表达最低,且该基因受ABA、干旱和高盐诱导上调表达。【结论】获得了BnbZIP1的全长cDNA序列,其编码蛋白具有植物bZIP转录因子典型的结构域,且该基因响应ABA、干旱和高盐逆境胁迫,预示该基因可能在植物抗逆反应中发挥着重要的调控作用。 [Objective] The objective of this study was to clone the full-length cDNA of BnbZIP1 transcription factor gene from ramie, and the expression pattern and the bioinformatics of the sequence were analyzed, and the prokaryotic expression and the subcellular localization were analyzed. [ Method] A full-length cDNA sequence was cloned by RT-PCR and RACE methods based on the unigene48047 in ramie transcriptome sequencing. Then the sequence was analyzed through bioinformatics methods and the expression patterns of BnbZIP1 were analyzed by using Real-time PCR in various tissues and under different stress conditions. A prokaryotic expression vector was constructed and the prokaryotic protein expression was induced with IPTG. Then a fusion expression vector containing EGFP was constructed to observe the subcellular localization. [Result]The full-length cDNA sequence and the ORF of BnbZIP1 were 2 071 bp and 1 407 bp, which encoded 468 amino acids with predicted pI and molecular weight were 4.95 and 36.81 kD, respectively. Homology comparison analysis showed that the deduced BnbZIPI amino acid sequence shares a 93% homology with bZIP gene (XP_002307972) in Populus trichocarpa. The relative molecular weight of recombinant protein induced by IPTG was 52 kD, which corresponded to the theoretical value. The results of subcellular localization analysis showed that the BnbZIP1 is located in nucleus. The results of real-time PCR suggested that the BnbZIP1 gene expressed in root, stem, shoot tipand blade, female flowers and male flowers, with the highest expression level in male flowers and the lowest in root. The BnbZIP1 gene was up-regulated by ABA, drought and high salt treatment. [ Conclusion ] The full-length cDNA sequence of BnbZIP1 from ramie was cloned and it had the typical bZIP transcription factor structural domain in plants, and BnbZIP1 gene responses to ABA, drought and high salt stress, which indicated that the BnbZIP1 genes might play an important role in stress response.
出处 《中国农业科学》 CAS CSCD 北大核心 2013年第7期1314-1322,共9页 Scientia Agricultura Sinica
基金 湖南省科技计划重点项目(2010TP4004-1 2010FJ2013 2012FJ4063) 国家科技支撑计划(2012BAD20B05-04)
关键词 苎麻 转录因子 BZIP 表达分析 亚细胞定位 ramie transcription factor bZIP expression analysis subcellular localization
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参考文献30

  • 1Farooq M, Wahid A, Kobayashi N, Fujita D, Basra S M A. Plant drought sa'ess: Effects, mechanisms and management. Agronomy for Sustainable Development, 2009, 29: 185-212.
  • 2周精华,邢虎成,揭雨成,钟英丽,朱守晶,蒋杰,王亮.苎麻Δ~1-吡咯啉-5-羧酸合成酶(P5CS)基因的克隆和表达分析[J].作物学报,2012,38(3):549-555. 被引量:13
  • 3张计育,渠慎春,郭忠仁,杜小丽,都贝贝,章镇.植物bZIP转录因子的生物学功能[J].西北植物学报,2011,31(5):1066-1075. 被引量:40
  • 4Chuang C F, Running M P, Williams R W, Meyerowitz E W. The PERIANTHIA gene encodes a bZIP protein involved in the determination of floral organ number in Arabidopsis thaliana. Genes and Development, 1999, 13: 334-344.
  • 5Iven T, Strathmann A, Botmer S, Zwafink T, Heinekamp T, Guivarc'h A, Roitsch T, Droge-Laser W. Homo-and heterodimers of tobacco bZIP proteins counteract as positive or negative regulators of transcription during pollen development. The Plant Journal, 2010, 63(1): 155-166.
  • 6Oflate L, Vieente-Carbajosa J, Lara P, Diaz I, Carbonero P. Barley BLZ2, a seed-specific bZIP protein that interacts with BLZI in vivo and activates transcription from the GCN4-1ike motif of B-hordein promoters in barley endosperm. The Journal of Biological Chemistry, 1999, 274: 9175-9182.
  • 7Onodera Y, Suzuki A, Wu C Y, Washida H, Takaiwa F. A rice functional transcriptional activator, RISBZI, responsible for endosperm-specific expression of storage protein genes through GCN4 motif. The Journal of Biological Chemistry, 2001, 276: 14139-14152.
  • 8Guo M, Chen Y, Du Y, Dong Y, Guo W, Zhai S, Zhang H, Dong S, Zhang Z, Wang Y, Wang P, Zheng X. The bZIP transcription factor MoAP1 mediates the oxidative stress response and is critical for pathogenicity of the dee blast fungus maguaporthe Oryzae. PLoS Pathogens, 2011, 7(2): el001302.
  • 9Lee S C, Choi H W, Hwang I S, Choi D S, Hwang B K. Functional roles of the pepper pathogen-induced bZIP transcription factor, C.4bZIP1, in enhanced resistance topathogen infection and environmental stresses. Planta, 2006, 224(5): 1209-1225.
  • 10Hsieh T H, Li C W, Su R C, Cheng C P, Sanjaya, Tsai Y C, Chan M T. A tomato bZIP transcription factor, SIAREB, is involved in water deficit and salt stress response. Planta, 2010, 231(6): 1459-1473.

二级参考文献162

  • 1VELASCO R,ZHARKIKH A,TROGGIO M,et al. A High quality draft consensus sequence of the genome of a heterozygous grapevine variety[J]. PLoS One, 2007,2(12) : 326.
  • 2MING R, HOU S,FENG Y,et al. The draft genome of the transgenic tropical fruit tree papaya (Carica papaya Linnaeus)[J]. Nature, 2008,452(7 190) :991--996.
  • 3PATERSON A H, BOWERS J E,BRUGGMANN R, et al. The Sorghum bicolor genome and the diversification of grasses[J]. Nature, 2009,457(7 229) :551--556.
  • 4SCHMUTZ J,CANNON S B,et al. Genome sequence of the palaeopolyploid soybean[J]. Nature,2010,463(7 278) :178--183.
  • 5VELASCO R, ZHARKIKH A, AFFOURTIT J ,et al. The genome of the domesticated apple (Malus)< domestica Borkh. )[J]. Nature Ge- netics, 2010,42(10) : 833 --839.
  • 6SCHWECHHEIMER C,et al. The regulation of transcription factor activity in plants[J].Trends Plant Sci. , 1998,3(10) :378--388.
  • 7LIU L,WHITE M J,et al. Transcription factors and their genes in higher plants[J]. Eur. J. Biochem. , 1999,262(2) :247--257.
  • 8EULGEM T, RUSHTON P J, ROBATZEK S, et al. The WRKY superfamily of plant transcription factors[J]. Trends Plant Sci. , 2000, 5(5) :199--206.
  • 9JAKOBY M, WEISSHAAR B, DROE LASER W, et al. The family of bZIP transcription factors in Arabidopsis thaliana [J]. Trends Plant Sci ,2002,7(3) :106--111.
  • 10DESPRES C, DELONG C, GLAZE S, et al. The Arabidopsis NPR 1/NIM1 protein enhances the DNA binding activity of a subgroup of the TGA family of bZIP transcription factors[J]. Plant Cell, 2000,12(2) :279--290.

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