大豆GmCOL1和GmCOL13启动子和基因组基因的克隆与生物信息学分析

doi:10.13560/J.cnki.biotech.bull.1985.2017-0201

摘要/Abstract

摘要： 以大豆天隆1号为材料提取基因组DNA，分别克隆到长2 631 bp的 GmCOL1启动子和2 809 bp的GmCOL13启动子。此外，还克隆到GmCOL1和GmCOL13基因组基因长度分别为3 488 bp和2 798 bp，它们分别编码含348个和366个氨基酸的蛋白质。利用PlantCARE分析GmCOL1和GmCOL13的启动子序列发现，它们均含有以下调控元件：生物钟元件（Circadian）、光响应元件（ACEG-box等）、ABA响应元件（ABRE）、干旱诱导元件（MBS）、低温响应元件（LTR）。ProtParam分析表明，GmCOL1和GmCOL13蛋白质的分子量分别为38.47 kD和40.91 kD，并且都是亲水性蛋白。GOR分析显示，GmCOL1和GmCOL13的二级结构含有Alpha helix，Extended strand和Random coil。在线网站PROSITE的分析说明，GmCOL1和GmCOL13有两个Zinc finger B-box和一个CCT domain（CO，CO-LIKE and TOC1 domain）结构域。

关键词: 大豆, GmCOL1, GmCOL13, 基因克隆, 生物信息学分析

Abstract: In the present study，the promoters and genomic genes of GmCOL1 and GmCOL13 were cloned from soybean cultivar Tianlong 1. The lengths of GmCOL1 and GmCOL13 promoters were 2 631 bp and 2 809 bp，while their genomic genes were 3 488 bp and 2 798 bp encoding 348 amino acids and 366 amino acids，respectively. Analysis by PlantCARE demonstrated that some important elements in the promoters of both GmCOL1 and GmCOL13：circadian，cis-acting regulatory element involved in light response，cis-acting element involved in ABA response，MYB binding site involved in drought-induction，and cis-acting element involved in low-temperature response. ProtParam analysis showed that GmCOL1 and GmCOL13 proteins were 38.47 kD and 40.91 kD，respectively，and both were hydrophilic. GOR analysis revealed that their secondary structure included mainly alpha helix，extended strand and random coil. Online PROSITE investigation displayed that GmCOL1 and GmCOL13 both had two zinc finger B-boxes and one CCT domain.

Key words: soybean, GmCOL1, GmCOL13, gene cloning, bioinformatics analysis

韩英英, 陈福禄, 傅永福, 张晓玫. 大豆^GmCOL1和^GmCOL13启动子和基因组基因的克隆与生物信息学分析[J]. 生物技术通报, 2017, 33(8): 63-72.

HAN Ying-ying, CHEN Fu-lu, FU Yong-Fu, ZHANG Xiao-mei. Cloning and Bioinformatics Analysis of Promoters and Genomic Genes of ^GmCOL1and ^GmCOL13 in Soybean[J]. Biotechnology Bulletin, 2017, 33(8): 63-72.

参考文献

[1] Komeda Y. Genetic regulation of time to flower in Arabidopsis thaliana[J] . Annu Rev Plant Biol, 2004, 55：521-535.
[2] Simpson GG, Dean C. Arabidopsis, the Rosetta stone of flowering time？[J] . Science, 2002, 296（5566）：285-289.
[3] Imaizumi T, Schultz TF, Harmon FG, et al. FKF1 F-box protein mediates cyclic degradation of a repressor of CONSTANS in Arabidopsis[J] . Science, 2005, 309（5732）：293-297.
[4] Kobayashi Y, Weigel D. Move on up, it’s time for change--mobile signals controlling photoperiod-dependent flowering[J] . Genes & Development, 2007, 21（19）：2371-2384.
[5] Turck F, Fornara F, Coupland G. Regulation and identity of florigen：FLOWERING LOCUS T moves center stage[J] . Annual Review of Plant Biology, 2008, 59：573-594.
[6] Song YH, Ito S, Imaizumi T. Flowering time regulation：photoperiod- and temperature-sensing in leaves[J] . Trends Plant Sci, 2013, 18（10）：575-583.
[7] Samach A, Onouchi H, Gold SE, et al. Distinct roles of CONSTANS target genes in reproductive development of Arabidopsis[J] . Science, 2000, 288（5471）：1613-1616.
[8] Koornneef M, Hanhart CJ, van der Veen JH. A genetic and physiological analysis of late flowering mutants in Arabidopsis thaliana[J] . Mol Gen Genet, 1991, 229（1）：57-66.
[9] Rédei GP. Supervital mutants of Arabidopsis[J] . Genetics, 1962, 47（4）：443-460.
[10] Sawa M, Nusinow DA, Kay SA, et al. FKF1 and GIGANTEA complex formation is required for day-length measurement in Arabidopsis[J] . Science, 2007, 318（5848）：261-265.
[11] Fornara F, Panigrahi KC, Gissot L, et al. Arabidopsis DOF transcription factors act redundantly to reduce CONSTANS expression and are essential for a photoperiodic flowering response[J] . Developmental Cell, 2009, 17（1）：75-86.
[12] Song YH, Smith RW, To BJ, et al. FKF1 conveys timing information for CONSTANS stabilization in photoperiodic flowering[J] . Science, 2012, 336（6084）：1045-1049.
[13] Valverde F, Mouradov A, Soppe W, et al. Photoreceptor regulation of CONSTANS protein in photoperiodic flowering[J] . Science, 2004, 303（5660）：1003-1006.
[14] Liu B, Zuo Z, Liu H, et al. Arabidopsis cryptochrome 1 interacts with SPA1 to suppress COP1 activity in response to blue light[J] . Genes & Development, 2011, 25（10）：1029-1034.
[15] Zuo Z, Liu H, Liu B, et al. Blue light-dependent interaction of CRY2 with SPA1 regulates COP1 activity and floral initiation in Arabidopsis[J] . Current Biology：CB, 2011, 21（10）：841-847.
[16] Laubinger S, Marchal V, Le Gourrierec J, et al. Arabidopsis SPA proteins regulate photoperiodic flowering and interact with the floral inducer CONSTANS to regulate its stability[J] . Development, 2006, 133（16）：3213-3222.
[17] Jang S, Marchal V, Panigrahi KC, et al. Arabidopsis COP1 shapes the temporal pattern of CO accumulation conferring a photoperiodic flowering response[J] . The EMBO Journal, 2008, 27（8）：1277-1288.
[18] Liu B, Kanazawa A, Matsumura H, et al. Genetic redundancy in soybean photoresponses associated with duplication of the phytochrome A gene[J] . Genetics, 2008, 180（2）：995-1007.
[19] Lazaro A, Valverde F, Pineiro M, et al. The Arabidopsis E3 ubiquitin ligase HOS1 negatively regulates CONSTANS abundance in the photoperiodic control of flowering[J] . The Plant Cell, 2012, 24（3）：982-999.
[20] Strizhov N, Abraham E, Okresz L, et al. Differential expression of two P5CS genes controlling proline accumulation during salt-stress requires ABA and is regulated by ABA1, ABI1 and AXR2 in Arabidopsis[J] . The Plant Journal, for Cell and Molecular Biology, 1997, 12（3）：557-569.
[21] Wang X, Fan C, Zhang X, et al. BioVector, a flexible system for gene specific-expression in plants[J] . BMC Plant Biol, 2013, 13（1）：198.
[22] Kim Y, Lim J, Yeom M, et al. ELF4 regulates GIGANTEA chromatin access through subnuclear sequestration[J] . Cell Rep 2013, 3（3）：671-677.
[23] Hotton SK, Callis J. Regulation of cullin RING ligases[J] . Annual Review of Plant Biology, 2008, 59：467-489.
[24] Fan C, Hu R, Zhang X, et al. Conserved CO-FT regulons contribute to the photoperiod flowering control in soybean[J] . BMC Plant Biology, 2014, 14：9.
[25] Ledger S, Strayer C, Ashton F, et al. Analysis of the function of two circadian-regulated CONSTANS-LIKE genes[J] . The Plant Journal, for Cell and Molecular Biology, 2001, 26（1）：15-22.
[26] Wong AC, Hecht VF, Picard K, et al. Isolation and functional analysis of CONSTANS-LIKE genes suggests that a central role for CONSTANS in flowering time control is not evolutionarily conserved in Medicago truncatula[J] . Frontiers in Plant Science, 2014, 5：486.
[27] González-Schain ND, Díaz-Mendoza M, Zurczak M, et al. Potato CONSTANS is involved in photoperiodic tuberization in a graft-transmissible manner[J] . The Plant Journal, for Cell and Molecular Biology, 2012, 70（4）：678-690.
[28] Sugiyama N, Nakagami H, Mochida K, et al. Large-scale phosphorylation mapping reveals the extent of tyrosine phosphorylation in Arabidopsis[J] . Molecular Systems Biology, 2008, 4：193.
[29] Sarid-Krebs L, Panigrahi KC, Fornara F, et al. Phosphorylation of CONSTANS and its COP1-dependent degradation during photoperiodic flowering of Arabidopsis[J] . The Plant Journal：for Cell and Molecular Biology, 2015, 84（3）：451-463.