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| cDNA Cloning, Bioinformatics Analysis and Construction of Overexpression Vector of High-chlorophyll Rice Gene DET1 |
| HUANG Jun-li1,LIU Tai-bo1,WANG Gui-xue1,LI Xian-yong1,2 |
1.College of Biological Engineering Chongqing University, Chongqing 400030, China
2.Rice Institute, Chongqing Academy of Agricultural Science，Chongqing 400060, China |
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Abstract The efficiency of photosynthesis can be improved by increasing chlorophyll content in rice leaves. In this study, DET1 （DE-ETIOLATED 1）gene related to high chlorophyll content has been cloned based on the fine mapping and sequence analysis of the DET1 gene was carried out in the high-chlorophyll mutants. The DET1 cDNA full-length of 1536 bp encoded 512 amino acids, and revealed a single T-to-C base transversion in the seventh exon of the DET1 gene which consisted of 11 exons, in comparison with the normal wild-type sequence. This transversion resulted in a conserved Leucine-to-Serine amino acid substitution at the position of 328, and introduced a recognition site for the EcoR I restriction endonuclease and the mutation located in the highly conserved region. Besides, the amino acid sequence of DET1 had more than 62% homology compared to corn and other higher plants. The phylogenetic tree, physical and chemical properties, hydrophilicity and hydrophobicity of the amino acid sequence have been analyzed by the bioinformatics technology. The overexpression vector of pCUPHN-DET1 has been constructed with the binary expression vector pCUPHN. The results laid good foundation for functional verification of DET1.
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Received: 30 December 2009
Published: 29 April 2010
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Corresponding Authors:
LIU TaiBo
E-mail: huang_junli@126.com;guixue_wang@126.com
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| ［1］ 李贤勇，王楚桃，李顺武，等.一个水稻高叶绿素含量基因的发现.西南农业学报，2002，15（4）: 122123. Li X Y, Wang C T, Li S W, et al. Southwest China Journal of Agricultural Sciences, 2002, 15（4）: 122123.
［2］ 李贤勇，王楚桃，李顺武，等.一个水稻高叶绿素基因的生物学特性研究.西南农业学报，2004，17（3）: 292294. Li X Y, Wang C T, Li S W, et al. Southwest China Journal of Agricultural Sciences, 2004, 17（3）: 292294.
［3］ Wang F H, Wang G X, Li X Y, et al. Heredity, physiology and mapping of a chlorophyll content gene of rice (Oryza sativa L.). Journal of Plant Physiology, 2008, 165(3):324330.
［4］ Mustilli A C, Fenzi F, Ciliento R, et al. Phenotype of the tomato high pigment2 mutant is caused by a mutation in the tomato homolog of DEETIOLATED1. Plant Cell, 1999, 11:145157.
［5］ Levin I, Frankel P, Gilboa N, et al. The tomato dark green mutation is a novel allele of the tomato homolog of the DEETIOLATED1 gene. Theor Appl Genet, 2003, 106:454460.
［6］ Pepper A, Delaney T, Washburn T, et al. DET1, a negative regulator of lightmediated development and gene expression in Arabidopsis, encodes a novel nuclearlocalized protein. Cell, 1994, 78:109116.
［7］ Song H R, Carre I A. DET1 regulates the proteasomal degradation of LHY, a component of the Arabidopsis circadian clock. Plant Molecular Biology, 2005, 57:761771.
［8］ Alexandrov N N, Brover W, Freidin S, et al. Insights into corn genes derived from largescale cDNA sequencing. Plant Mol Biol, 2009, 69:179194.
［9］ Paterson A H, Bowers J E, Bruggmann R, et al. The Sorghum bicolor genome and the diversification of grasses. Nature, 2009, 457 (7229):551556.
［10］ 范敏，金黎平，刘庆昌，等.马铃薯抗旱机理及其相关研究进展.中国马铃薯，2006，32（2）: 101107. Fan M, Jin L P, Liu Q C, et al. Chinese Potato, 2006, 32(2):101107.
［11］ Qiu D W, Mao J J, Yang X F, et al. Expression of an elicitorencoding gene from Magnaporthe grisea enhances resistance against blast disease in transgenic rice. Plant Cell Report, 2008, 28:925933. |
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