DELETION ANALYSIS OF A 2S SEED STORAGE PROTEIN PROMOTER OF BRASSICA-NAPUS IN TRANSGENIC TOBACCO

被引:89
作者
STALBERG, K
ELLERSTROM, M
JOSEFSSON, LG
RASK, L
机构
[1] Uppsala Genetic Center, Department of Cell Research, Swedish University of Agricultural Sciences, Uppsala, S-750 07
关键词
BRASSICA NAPUS; DELETION ANALYSIS; NAPIN; PROMOTER; SEED; STORAGE PROTEIN;
D O I
10.1007/BF00021523
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
The promoter and upstream region of the Brassica napus 2S storage protein napA gene were studied to identify cis-acting sequences involved in developmental seed-specific expression. Fragments generated by successive deletions of the 5' control region of the napA gene were fused to the reporter gene beta-glucuronidase (GUS). These constructs were used to transform tobacco leaf discs. Analyses of GUS activities in mature seeds from the transformed plants indicated that there were both negatively and positively acting sequences in the napin gene promoter. Deletion of sequences between -1101 and -309 resulted in increased GUS activity. In contrast, deletion of sequences between -309 and -211 decreased the expression. The minimum sequence required for seed-specific expression was a 196 bp fragment between -152 and +44. Further 5' deletion of the fragment to -126 abolished this activity. Sequence comparison showed that a G box-like sequence and two sequence motifs conserved between 2S storage protein genes are located between -148 to -120. Histochemical and fluorometric analysis of tobacco seeds showed that the spatial and developmental expression pattern was retained in the deletion fragments down to -152. However, the expression in tobacco seeds differed from the spatial and temporal expression in B. napus. In tobacco, the napA promoter directed GUS activity early in the endosperm before any visible activity could be seen in the heart-shaped embryo. Later, during the transition from heart to torpedo stages, the main expression of GUS was localized to the embryo. No significant GUS activity was found in either root or leaf.
引用
收藏
页码:671 / 683
页数:13
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