Volume 4, Issue 1, March 2019, Page: 7-11
Cloning and Expression Analysis of GmCYP78A5 Promoter
Xiaofeng Chen, College of Life Sciences, Qingdao Agricultural University, Qingdao, China; Sales Department, Qingdao Betterpet Foodstuff Company, Qingdao, China
Qiuli Du, Quancheng College, University of Jinan, Penglai, China
Chunmei Zhao, College of Life Sciences, Qingdao Agricultural University, Qingdao, China
Zhaoyong Lv, College of Life Sciences, Qingdao Agricultural University, Qingdao, China
Ren-Gao Xue, College of Life Sciences, Qingdao Agricultural University, Qingdao, China
Received: Apr. 21, 2019;       Accepted: Jun. 17, 2019;       Published: Jul. 4, 2019
DOI: 10.11648/j.ajpb.20190401.12      View  21      Downloads  5
Abstract
CYP78A5 promoter was isolated from soybean (Glycine max L. Merrill) plant by using PCR technology. DNA sequence alignment indicated that the amplified fragment (1650bp) was 99.21% homologous to the correspondent regions of the reported sequences. Bioinformatics analysis showed that the GmCYP78A5 promoter contains a lot of inducible or tissue-specific expression elements. RT-PCR results indicated that the gene GmCYP78A5 highly expressed in immature seed, weekly expressed in stem of soybean, but no expressed in root, leaf and flower. To further study the tissue expression patterns of GmCYP78A5 gene, the promoter of the gene GmCYP78A5 was fused with GUS reporter gene to construct a plant expression vector and the vector was transformed into tobacco (Nicotiana tabacum) by Agrobacterium-meditated method. The expression of the GUS gene in the transgenic tobacco plants indicated that the GmCYP78A5 promoter could drive the GUS reporter gene to express highly in the leaf, stem, sepal, pedicel, seeds of the transgenic tobacco plants, demonstrating that the expression patterns of the GmCYP78A5 promoters in soybean and tobacco were inconsistent.
Keywords
Soybean, GmCYP78A5 Promoter, Tissue Specific Expression, GUS Assay
To cite this article
Xiaofeng Chen, Qiuli Du, Chunmei Zhao, Zhaoyong Lv, Ren-Gao Xue, Cloning and Expression Analysis of GmCYP78A5 Promoter, American Journal of Plant Biology. Vol. 4, No. 1, 2019, pp. 7-11. doi: 10.11648/j.ajpb.20190401.12
Copyright
Copyright © 2019 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Reference
[1]
Wang X, Li Y, Zhang H, Sun G, Zhang W, and Qiu L (2015). Evolution and association analysis of GmCYP78A10 gene with seed size/weight and pod number in soybean. Mol Biol Rep, 42:489-496.
[2]
Anastasiou E, Kenz S, Gerstung M, MacLean D, Timmer J, Fleck C, and Lenhard M (2007). Control of Plant Organ Size by KLUH/CYP78A5-Dependent Intercellular Signaling. Developmental Cell, 13: 843-856.
[3]
Eriksson S, Stransfeld L, Adamski N M, Breuninger H, Lenhard M (2010). Local maternal control of seed size by KLUH/CYP78A5-dependent growth signaling. Curr Biol, 20 (6):527-32.
[4]
Zondlo S C and Irish V F (1999). CYP78A5 encodes a cyptochrome P450 that marks the shoot apical meristem boundary in Arabidopsis. Plant J, 19 (3): 259-268.
[5]
Anastasiou E, and Lenhard M (2007). Growing up to one's standard. Curr. Opin. Plant Biol, 10 (1): 63-69.
[6]
Adamski N M, Anastasiou E, Eriksson S, O'Neill C M, and Lenhard M (2009). Local maternal control of seed size by KLUH/CYP78A5-dependent growth signaling. Proc. Natl. Acad. Sci. USA, 106 (47): 20115-20120.
[7]
Zhong M, Huang G. T, Bai L. P, Zhang L, Ma H, Zhang L. J, and Guo Z. H (2011). New strategy to reconstruct Agrobacterium-mediated plant transgenic expression vectors. Acta Agriculturae Boreali Sinica, 26 (1): 41-46.
[8]
Wei J, Mao W H., Lin Y J, and Chen H (2012). Isolation and characterization of a novel rice constitutive promoter. Journal of Huazhong Agricultural University, 31 (2): 139-146.
[9]
Li H, and Wang X L (1999) The difficulties in the isolation of RNA from plant tissues and their resolving strategies. Biotechnology Information, 1: 36-39.
[10]
Zhao SY, Wu Y R, and Xia G. M (2002). Introduction of a simple and effective method for plant total RNA isolation. Hereditas, 24 (3): 337-338.
[11]
Du D L, Ma W R, Su J, Zhou P, and Zheng X Q (2003). A comparative study of genomic DNAS from banana extracted by SDS, CTAB and PVP methods. Journal of Hainan Normal University (Natural Science), 16 (1):74-80.
[12]
Zheng W J, Liu X, Liu W, Tang D Z, He L, and Zhang H W (2003). Qualitative analysis of the processed genetically modified soybean products by PCR-based methods. Journal of Agricultural Biotechnology, 11 (5): 467-471.
[13]
Hofen R, and Willmitzer L (1988). Storage of competent cells for Agrobactertium transformation. Nucleic Acids Res, 16: 9877.
[14]
Jefferson R A (1987). Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol. Biol. Rep, 5: 384-405.
[15]
Hou B K, Xia G M, and Chen Z H (2001). Strategies for optimizing expression vectors used in plant genetic engineering. Hereditas, 23 (5): 492-495.
[16]
Zhang C X, Wang W Q, Jiang X N, and Chen X M (2004). Review on plant gene promoters. Acta Genetica Sinica, 31 (12): 1455-1464.
[17]
LV Z, Zhao C, and Xue R (2016). Cloning and expression analysis of grape’s stress inducible promoter. Acta Agriculturae Boreali-Sinica, 31 (1): 77-82.
[18]
Zhao C, Fan X, and Xue R (2017). Molecular cloning and function analysis of a GmCHI1 Promoter. Acta Agriculturae Boreali-Sinica, 32 (4): 32-36.
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