Isolation and analysis of 5' regulatory sequence of VVICOR11 from cowberry
2012
Wang, Q.J. | Gao, Z.H. | Tao, J.M. | Qu, S.C. | Zhang, Z.
The gene of vvicor11 isolated from cowberry is a KS type dehydrin gene, which belongs to LEA D11 or LEA II protein family. LEA (late embryogenesis-abundant protein) proteins accumulate during dehydration, low temperature, freezing, abscisic acid (ABA) treatment, and seed maturation. To better understand its expression and regulation, we isolated an 1110 bp 5’ region upstream starting site of the putative vvicor11 (GenBank No. FJ429387). The sequence was identified using a genome walking strategy based on the cDNA sequence of cowberry vvicor11. Computational analysis using the PLACE and PlantCARE databases showed that the sequence included 5’-UTR Pyrimidine-rich motifs and typical promoter characterization structures, such as a TATA-box and a CAAT-box, to ensure the proper transcription initiation. There were many potential cis-acting elements involved in stress, such as ABRE, LTR, TC-rich repeats, ARE, MYB (MYC), TGACG-motif, CBFHV, and so on. The presence of these motifs could suggest that the expression of vvicor11 could be induced by ABA, low temperature, drought and high salt stress, and could be a downstream target gene of the CBF transcription factor. Transient expression of a 5’- regulatory sequence deletion series showed that the degree of 5’- regulatory sequence activation was increased the longer the sequences. One of the constructions, the CP5::GUS construct was expressed in the root, stem, leaf, petal and style of tobacco without inductive treatment. This suggested that the 5’- regulatory sequence possessed at least basal expression in various organs of a different species.
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