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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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