全部 标题 作者
关键词 摘要

OALib Journal期刊
ISSN: 2333-9721
费用:99美元

查看量下载量

相关文章

更多...
大豆科学  2015 

大豆GmbZIP基因的克隆及EMSA分析

DOI: 10.11861/j.issn.1000-9841.2015.01.0032

Keywords: 大豆,bZIP,原核表达,EMSA

Full-Text   Cite this paper   Add to My Lib

Abstract:

通过PCR扩增的方法从大豆品种Harosoy中克隆到1个新的bZIP基因(GmbZIP71),基因表达分析表明GmbZIP71在叶片、生长点、花、花芽、荚和根等多个器官中表达。将GmbZIP71构建到原核表达载体pET29b上,导入大肠杆菌BL21(DE3)中,对其进行IPTG诱导。结果表明:在IPTG浓度为0.25mmol?L1,诱导时间为4h,诱导温度为28℃时,重组蛋白得到表达,分子量大约为48kDa,SDS-PAGE电泳结果表明重组蛋白主要以包涵体形式存在,用His蛋白纯化系统回收得到GmbZIP71-His重组蛋白,EMSA(electrophoreticmobilityshiftassay)实验表明GmbZIP71重组蛋白可以在体外与ACGT顺式作用元件结合。

References

[1]  [1]Glover J M, Harrison S C. Crystal structure of the heterodimeric bZIP transcription factor c-Fos-c-Jun bound to DNA[J]. Nature, 1995, 373: 257-261.
[2]  [2]Jakoby M, Weisshaar B, Dr o..?ge-Laser W, et al. bZIP transcription factors in Arabidopsis [J]. Trends in Plant Science, 2002, 7(3): 106-111.
[3]  [3]Liao Y, Zhang J S, Chen S Y, et al. Role of soybean GmbZIP132 under abscisic acid and salt stresses[J]. Journal of Integrative Plant Biology, 2008, 50(2): 221-230.
[4]  [4]Liao Y, Zou H F, Wei W, et al. Soybean GmbZIP44, GmbZIP62and GmbZIP78 genes function as negative regulator of ABA signaling and confer salt and freezing tolerance in transgenic Arabidopsis[J]. Planta, 2008, 228(2): 225-240.
[5]  [5]Gao S Q, Chen M, Xu Z S, et al. The soybean GmbZIP transcription factor enhances multiple abiotic stress tolerances in transgenic plants[J]. Plant Molecular Biology, 2011, 75(6): 537-553.
[6]  [6]才华, 朱延明, 柏锡, 等. 野生大豆GsbZIP33基因的分离及胁迫耐性分析[J]. 分子植物育种, 2011, 9(4): 397-401.(Cai H, Zhu Y M, Bai X, et al. Isolation and tolerance analysis of GsbZIP33 gene linked to response on stress in Glycine soja[J]. Molecular Plant Breeding, 2011, 9(4): 397-401)
[7]  [7]Xu L, Xu Z, Liu X, et al. The subcellular localization and ectopic expression analysis in Arabidopsis of soybean GmbZIP60 gene[J]. Journal of Plant Biochemistry and Biotechnology, 2013: 10.1007/s13562-013-0228-4.
[8]  [8]Foster R, Izawa T, Chua N H. Plant bZIP proteins gather at ACGT elements[J]. The FASEB Journal, 1994, 8(2): 192-200.
[9]  [9]Izawa T, Foster R, Chua N H. Plant bZIP protein-DNA binding-specificity[J]. Journal of Molecular Biology, 1993, 230(4): 1131-1144.
[10]  [10]Martinez-Garcia J F, Moyano E, Alcocer M J C, et al. Two bZIP proteins from Antirrhinum flowers preferentially bind a hybrid C-box/G-box motif and help to define a new subfamily of bZIP transcription factors[J]. The Plant Journal, 1998, 13(4): 489-505.
[11]  [11]Xiang C, Miao Z, Lam E. DNA-binding properties, genomic organization and expression pattern of TGA6, a new member of the TGA family of bZIP transcription factors in Arabidopsis thaliana[J]. Plant Molecular Biology, 1997, 34: 3-15.
[12]  [12]Grigoryan G, Keating A E. Structure-based prediction of bZIP partnering specificity[J]. Journal Molecular Biology, 2006, 355(5): 25-42.
[13]  [13]Terzaghi W B, Bertekap R L, Cashmore A R. Intracellular localization of GBF proteins and blue light-induced import of GBF2 fusion proteins into the nucleus of cultured Arabidopsis and soybean cells[J]. The Plant Journal, 1997, 11(5): 67-82.
[14]  [14]Riechmann J L, Heard J, Martin G, et al. Arabidopsis ?transcription factors:Genome wide comparative analysis among eukaryotes[J]. Science, 2000, 290: 2105-2110.
[15]  [15]Paulo S S, Fábio T S, Nogueira, et al. Identification of new ABA- and MEJA-activated sugarcane bZIP genes by data mining in the SUCEST database[J]. Plant Cell Report, 2008, 27: 335-345.
[16]  [16]Guiltinan M J, Marcottel A. Plant leucine zipper protein that recognizes an abscisic and response element[J] . Science, 1990, 250: 267-270.
[17]  [17]Choi H, Hong J, Kang J, et al. ABFs, a family of ABA responsive element binding factors[J] . The Journal of Biological Chemistry, 2000, 21: 1723-1730.
[18]  [18]Fujita Y, Fujita M, Satoh R, et al. AREB1 is a transcription activator of novel ABRE2 dependent ABA signaling that enhances drought stress tolerance in Arabidopsis[J]. The Plant Cell, 2005, 17: 3470-3488.
[19]  [19]Kang J, Choi H, Kim S Y. Arabidopsis basic leucine zipper proteins that mediate stress responsive abscisic acid signaling[J]. The Plant Cell, 2002,14: 343-357.
[20]  [20]Kim J B, Kang J Y, Kim S Y. Over-expression of a transcription factor regulating ABA responsive gene expression confers multiple stress tolerance[J]. Plant Biotechnology Journal, 2004, 2(5): 459-466.

Full-Text

comments powered by Disqus

Contact Us

service@oalib.com

QQ:3279437679

WhatsApp +8615387084133