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| Cloning and Activity Analysis of Soybean FAD2-1B Promoter |
| ZHAO Yan1, SHA Wei1, ZHANG Mei-juan1, YANG Xiao-jie1, FAN Zhen-yu1, WANG Yan-mei2 |
1. College of Life Science and Agroforestry, Qiqihaer University, Qiqihaer 161000, China;
2. LongSha Park of Qigihar City, Qiqihaer 161006, China |
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Abstract The soybean oleate desaturase gene(FAD2-1B ) has previously been shown to be expressed specifically in soybean seeds. The 5'-flanking upstream sequence of FAD2-1B gene, named FP, was isolated from the genomic DNA of soybean by PCR method. Sequence analysis by PLACE revealed that this fragment contains a series of motifs related to seed-specific promoters, such as Skn-1 motif,AACACA,SEF4 motif,E-box,ACGT. Replacing CaMV35S promoter of pCAMBIA1301 with FP fragment, the binary expression vector pCAM-FP was constructed. Transient expression in soybean tissues by Agrobacterium tumefaciens mediated method, the results of histochemical GUS analysis showed that there were little or not GUS activities in roots, stems and leaves, but there was higher activity in soybean seeds. It is inferred that FP promoter possess the function driven downstream gene expression exclusively in soybean seeds.
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Received: 12 October 2012
Published: 25 April 2013
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[1] 肖钢, 张宏军, 彭琪等. 甘蓝型油菜油酸脱氢酶基因(fad2)多个拷贝的发现及分析. 作物学报, 2008, 34(9): 1563-1568. Xiao G, Zhang H J, Peng Q, et al. Screening and analysis of mutiple copy of oleate desaturase gene (fad2) in Brassica napus.Acta Agronomica Sinica,2008, 34(9): 1563-1568.
[2] Chen W, Song K, Cai Y R, et al. Genetic modification of soybean with a novel grafting technique: downregulating the FAD2-1 gene increases oleic acid content. Plant Mol Biol Rep, 2011, 29: 866-874.
[3] Sandhu D, Alt J L, Scherder C W, et al. Enhanced oleic acid content in the soybean mutant M23 is associated with the deletion in the Fad2-1a gene encoding a fatty acid desaturase. J Amer Oil Chem Soc, 2007, 84(3): 229-235.
[4] Chi X Y, Yang Q L, Pan L J, et al. Isolation and characterization of fatty acid desaturase genes from peanut (Arachis hypogaea L.). Plant Cell Rep, 2011, 30(8): 1393-1404.
[5] Jung J H, Kim H J, Go Y S, et al. Identi?cation of functional BrFAD2-1 gene encoding microsomal delta-12 fatty acid desaturase from Brassica rapa and development of Brassica napus containing high oleic acid contents. Plant Cell Rep, 2011, 30(10): 1881-1892.
[6] Li L Y, Wang X L, Gai J Y, et al. Molecular cloning and characterization of a novel microsomal oleate desaturase gene from soybean. J Plant Physiol, 2007, 164(11): 1516-1526.
[7] Li L, Wang X, Gai J, et al. Isolation and characterization of a seed-specific isoform of microsomal omega-6 fatty acid desaturase gene (FAD2-1B) from soybean. DNA Sequence, 2008, 19(1): 28-36.
[8] Xiao Y H, Luo M, Fang W G, et al. PCR walking in cotton genome using YADE method. Acta Genet Sin, 2002, 29(1): 62-66.
[9] Hu X W, Liu S X, Guo J C, et al. Embryo and anther regulation of the mabinlin Ⅱ sweet protein gene in Capparis masaikai Levl. Funct Integr Genomics, 2009, 9(3): 351-361.
[10] Jefferson R A. Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep, 1987, 5 (4): 387-405.
[11] Tamar J G, Hsu J Y, Theisen J W, et al. The RNA polymerase II core promoter--the gateway to transcription. Curr Opin Cell Biol, 2008, 20(3): 253-259.
[12] Wu C Y, Washida H, Onodera Y, et al. Quantitative nature of the Prolamin-box, ACGT and AACA motifs in a rice glutelin gene promoter: Minimal cis-element requirements for endosperm-specific gene expression. Plant J, 2000, 23(3): 415-421.
[13] Fauteux F, Stromvik M V. Seed storage protein gene promoters contain conserved DNA motifs in Brassicaceae, Fabaceae and Poaceae. BMC Plant Biol, 2009, 9:126.
[14] Kim M J, Kim J K, Shin J S, et al. The SebHLH transcription factor mediates trans-activation of the SeFAD2 gene promoter through binding to E-and G-box elementsv. Plant Mol Biol, 2007, 64 (4): 453-466.
[15] Chung K J, Hwang S K, Hahn B S, et al. Authentic seed-specific activity of the Perilla oleosin 19 gene promoter in transgenic Arabidopsis. Plant Cell Rep, 2008, 27(1): 29-37.
[16] Luo K, Zhang G F, Deng W, et al. Functional characterization of a cotton late embryogenesis-abundant D113 gene promoter in transgenic tobacco. Plant Cell Rep, 2008, 27(4):707-717.
[17] Gao M J, Lydiate D J, Li X, et al. Repression of seed maturation genes by a trihelix transcriptional repressor in Arabidopsis seedlings. The Plant Cell, 2009, 21(1):54-71.
[18] Chamberland S, Daigle N, Bernier F. The legumin boxes and the 3' part of a soybean beta-conglycinin promoter are involved in seed gene expression in transgenic tobacco plants. Plant Mol Biol, 1992, 19 (6): 937-949.
[19] Nagano Y, Furuhashi H, Inaba T, et al. A novel class of plant-specific zinc-dependent DNA-binding protein that binds to A/T-rich DNA sequences. Nucheic Acids Res, 2001, 29(20): 4097-4105.
[20] 李吉涛, 郭建春. 种子特异性启动子的研究进展. 安徽农业科学, 2008, 36(4): 1382-1385. Li J T, Guo J C. Advances in studies of seed-specific promoters.Journal of Anhui Agri Sci, 2008, 36(4): 1382-1385.
[21] Zhang Y J, Li L, Song Y R. Identification of seed-specifc promoter nap300 and its comparison with 7S promoter. Prog Nat Sci, 2002, 12(10): 737-741.
[22] Wu C Y, Suzuki A, Washida H, et al. The GCN4 motif in a rice glutelin gene is essential for endosperm-specific gene expression and is activated by Opaque-2 in transgenic rice plants. Plant J, 1998,14(6):673-683. |
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