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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志  2011, Vol. 31 Issue (5): 42-47    
研究报告     
毛白杨PtSEP2基因启动子克隆和瞬时表达特性分析
崔东清, 叶梅霞, 刘军梅, 李昊, 张志毅, 安新民
北京林业大学林木育种国家工程实验室 林木花卉遗传育种教育部重点实验室 国家林业局树木花卉育种与生物工程重点开放实验室 北京 100083
Cloning and Transient Expression Analysis of PtSEP2 Promoter from Populus tomentosa
CUI Dong-qing, YE Mei-xia, LIU Jun-mei, LI Hao, ZHANG Zhi-yi, AN Xin-min
National Engineering Laboratory for Tree Breeding; Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education; The Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration, Beijing Forestry University, Beijing 100083, China
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摘要:

利用PCR 技术从毛白杨基因组DNA中扩增获得花器官发育相关的 SEPALLATA2 类似基因PtSEP2 5'侧翼约2.3kb的一段序列,经PlantCARE序列分析表明,该序列中含有启动子特征的保守序列及多种光应答元件,初步推测其为 PtSEP2 基因启动子。进一步以GUS为报告基因,构建了 pPtSEP2 promoter∷GUS的植物表达载体, 命名为 PtSEP2p∷GUS 。以烟草根、茎、叶、花芽为受体,通过农杆菌介导转 化植物表达载体 PtSEP2p∷GUS 进行瞬时表达研究,结果表明该启动子能够驱动GUS报告基因在烟草花药中表达,而其表达活性弱于组成型表达的花椰菜花叶病毒(CaMV)35S启动子。该启动子的获得对于杨树及其他开花调控基因工程研究具有重要的应用价值。

关键词: 毛白杨SEPALLATA2启动子GUS活性瞬时表达    
Abstract:

A 5' flanking sequence of PtSEP2 which is a SEPALLATA2 -like gene involved in floral organ development was amplified by PCR from the genomic DNA of Populus tomentosa. The length of fragment is approximately 2.3kb. The results derived from PlantCARE analysis showed that the sequence contains conserved cis-acting elements of promoter, coupled with a variety of light-responsive elements. Therefore, it was speculated preliminarily to be the promoter of PtSEP2 gene. To investigate function of this promoter, it was fused to GUS reporter gene, generating a plant expression vector pPtSEP2 promoter∷GUS, named PtSEP2p∷GUS . With roots, stems, leaves and flower buds of Nicotiana tobaccum for the receptor, the results of transient expression that Agrobacterium-mediated that showed the PtSEP2 promoter was competent to drive specially GUS reporter gene expression in anther, yet its’ activity was weaker than that of cauliflower mosaic virus (CaMV) 35S promoter, which constitutively expressed in planta. Consequently, provides a possible genetic modification tool for flowering regulation in poplar and other plants.

Key words: Populus tomentosa    SEPALLATA2 promoter    GUS activity    Transient expression
收稿日期: 2010-11-24 出版日期: 2011-05-27
ZTFLH:  Q786  
基金资助:

国家自然科学基金(30571511)、国家林业公益性项目(201004009)、教育部重点项目(108017)资助项目

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引用本文:

崔东清, 叶梅霞, 刘军梅, 李昊, 张志毅, 安新民. 毛白杨PtSEP2基因启动子克隆和瞬时表达特性分析[J]. 中国生物工程杂志, 2011, 31(5): 42-47.

CUI Dong-qing, YE Mei-xia, LIU Jun-mei, LI Hao, ZHANG Zhi-yi, AN Xin-min. Cloning and Transient Expression Analysis of PtSEP2 Promoter from Populus tomentosa. China Biotechnology, 2011, 31(5): 42-47.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/        https://manu60.magtech.com.cn/biotech/CN/Y2011/V31/I5/42


[1] 张春晓, 王文棋, 蒋湘宁, 等. 植物基因启动子研究进展. 遗传学报, 2004, 31(12): 1455-1464. Zhang C X, Wang W Q, Jiang X N, et al. Journal of Genetics and Genomics, 2004, 31(12): 1455-1464.

[2] 侯丙凯, 夏光敏, 陈正华. 植物基因工程表达载体的改进和优化策略. 遗传, 2001,23(5): 492-497. Hou B K, Xia G M, Chen Z H. Hereditas (Beijing), 2001, 23(5): 492-497.

[3] Potenza C, Aleman L, Sengupta-Gopalan C. Targeting transgene expression in research, agricultural, and environmental applications: promoters used in plant transformation. In Vitro Cellular & Developmental BiologyPlant, 2004,40(1): 1-22.

[4] Wei H, Meilan R, Brunner A M, et al. Transgenic sterility in Populus: expression properties of the poplar PTLF, Agrobacterium NOS and two minimal 35S promoters in vegetative tissues. Tree Physiology, 2006, 26(4): 401-410.

[5] 王正加. 山核桃分子标记与开花基因CcLFY及其启动子克隆的研究.北京:北京林业大学,生物科学与技术学院, 2006. Wang Z J. Molecular Marker Analysis and Flora Gene CcLFY and Its Promoter Cloning of Carya Cathayensis. Beijing: Beijing Forestry University, College of Biological Sciences and Technology, 2006.

[6] 陈聪. 茶树花特异表达启动子-花粉壁蛋白基因启动子表达研究. 安徽: 安徽农业大学, 生命科学学院, 2009. Chen C. Study on Cpcp (Pollen Coat Protein Gene) Promoter Expression of Tea Plant (Camellia Sinensis). Anhui : Anhui Agricultural University, College of Life Science, 2009.

[7] 王静澄, 李昊, 崔东清, 等. 毛白杨 PtSEP3-1 基因启动子的克隆分析及其表达载体构建. 基因组学与应用生物学, 2010, 29(2): 239-244. Wang J C, Li H, Cui D Q, et al. Genomics and Applied Biology, 2010, 29(2): 239-244.

[8] An G. Binary Ti vectors for plant transformation and promoter analysis. Methods in enzymology, 1987, 153: 292-305.

[9] Jefferson R A, Kavanagh T A, Bevan M W. GUS fusions: beta-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J, 1987, 6(13): 3901-3907.

[10] Shahmuradov L A, Solovyev V V, Gammerman A J. Plant promoter prediction with confidence estimation. Nucleic Acids Research, 2005, 33(3): 1069-1076.

[11] Higo K, Ugawa Y, Iwamoto M, et al. Plant cis-acting regulatory DNA elements (PLACE) database: 1999. Nucleic Acids Research, 1999, 27(1): 297-300.

[12] Lescot M, Déhais P, Thijs G, et al. Plant-CARE: a database of plant cis-acting regulatory elements and a portal to tools for in silico analysis of promoter sequences. Nucleic Acids Research, 2002, 30(1): 325-327.

[13] Zheng H Q, Lin S Z, Zhang Q, et al. Isolation and analysis of a TIR-specific promoter from poplar. Forestry Studies in China, 2007, 9(2): 95-106.

[14] Jansson S, Douglas C J. Populus: a model system for plant biology. Annu Rev Plant Biol, 2007, 58:435-458.

[15] Leebens-Mack J, Soltis D E, Soltis P S. Plant reproductive genomics at the plant and animal genome conference. Comp Funct Genomics, 2005, 6(3): 159-169.

[16] Castillejo C, Romera-Branchat M, Pelaz S. A new role of the Arabidopsis SEPALLATA3 gene revealed by its constitutive expression. The Plant Journal, 2005, 43(4): 586-596.

[17] Rainer M, Günter T. Reconstitution of "floral quartets" in vitro involving class B and class E floral homeotic proteins. Nucleic Acids Res, 2009, 37(8): 2723-2736.

[18] Mizukami Y, Ma H. Ectopic expression of the floral homeotic gene AGAMOUS in transgenic Arabidopsis plants alters floral organ identity. Cell, 1992, 7l (1): 119-131.

[19] Cseke L J, Cseke S B, Ravinder N, et al. SEP-class genes in Populus tremuloides and their likely role in reproductive survival of poplar trees. Gene, 2005, 358: 1-16.

[20] Hua X J, van de Cotte B, van Montagu M, et al. The 5' untranslated region of the At-P5R gene is involved in both transcriptional and post-transcriptional regulation. Plant J, 2001, 26(2): 157-169.

[21] Kim M J, Kim H, Shin J S, et al. Seed-specific expression of sesame microsomal oleic acid desaturase is controlled by combinatorial properties between negative cis-regulatory elements in the SeFAD2 promoter and enhancers in the 5'-UTR intron. Mol Genet Genomics, 2006, 276(4): 351-368.

[22] Chung B Y, Simons C, Firth A E, et al. Effect of 5'-UTR introns on gene expression in Arabidopsis thaliana. BMC Genomics, 2006,7(5): 120.

[23] Samadder P, Sivamani E, Lu J L, et al. Transcriptional and post-transcriptional enhancement of gene expression by the 5' UTR intron of rice rubi3 gene in transgenic rice cells. Mol Genet Genomics, 2008, 79(4): 429-439.

[24] Karthikeyan A S, Ballachanda D N, Raghothama K G. Promoter deletion analysis elucidates the role of cis elements and 5' UTR intron in spatiotemporal regulation of AtPht1;4 expression in Arabidopsis. Physiol Plant, 2009, 136 (1): 10-18.

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