Articles
ISOLATION AND ANALYSIS OF 5′ REGULATORY SEQUENCE OF VVICOR11 FROM COWBERRY
Article number
926_14
Pages
107 – 111
Language
English
Abstract
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.
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.
Authors
Q.J. Wang, Z.H. Gao, J.M. Tao, S.C. Qu, Z. Zhang
Keywords
Vaccinium vitis-idaea, 5′ regulatory sequence of vvicor11, Agrobacterium– mediated transformations, tobacco, transient expression
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