Cloning and Characterization of the LEA3 Protein Gene and Promoter from Sorghum bicolor

doi:10.7677/ynzwyj201312110

Abstract

Abstract:

A pair of degenerate primers was designed according to reported fragments of LEA3 genes in GenBank. Using these primers a new cDNA fragment of 1032bp was amplified by PCR and RACE from total RNA of Sorghum bicolor, which indicated that the open reading frame region of the gene was 612bp (GenBank accession number GQ494000). It is predicted that the sequence encodes 203 amino acids with a molecular weight of about 21.22kD, includes seven characteristic motifs of LEA3 and has a theoretical isoelectric point of 8.79. The sequence shows a similarity of 73.8%, 53.77%, 45.63% and 53.99% with Zea mays, Triticum aestivum, Oryza sativa and Hordeum vulgare LEA3 sequences, respectively. The secondary structure of the Sorghum LEA3 protein exhibited a quantity of αhelix dominant structures that were similar to those found in many other plants. In addition, a 749bp DNA sequence was isolated from Sorghum genomic DNA by Thermal Asymmetric Interlaced PCR (TailPCR). This promoter region was predicted to contain ABA and drought stress response elements of embryo and endospermspecific genes Sequences of Gramineae LEA3 genes were aligned using the profile alignment function of ClustalX, and maximum likelihood phylogenetic analyses were conducted using PHyML. The research forms part of an analysis of droughtresistant mechanisms and gene function in Sorghum.

Key words: Sorghum bicolor, LEA3 protein, Clone, Promoter, ML tree

CLC Number:

Q 78

SUN Xiao-Jiao, SHANG Xiao-Qian, XU Li-Xia, BAI Qiong, YAN Bo. Cloning and Characterization of the LEA3 Protein Gene and Promoter from Sorghum bicolor[J]. Plant Diversity, 2013, 35(5): 585-593.

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