Sequences responsible for the tissue specific promoter activity of a pea legumin gene in tobacco.
Maturing pea cotyledons accumulate large quantities of storage proteins at a specific time in seed development. To examine the sequences responsible for this regulated expression, a series of deletion mutants of the legA major seed storage protein gene were made and transferred to tobacco using the Bin19 disarmed Agrobacterium vector system. A promoter sequence of 97 bp including the CAAT and TATA boxes was insufficient for expression. Expression was first detected in a construct with 549 bp of upstream flanking sequence which contained the the leg box element, a 28 bp conserved sequence found in the legumin-type genes of several legume species. Constructs containing -833 and -1203 bp of promoter sequence significantly increased levels of expression. All expressing constructs preserved seed specificity and temporal regulation. The results indicate that promoter sequences between positions -97 and -549 bp are responsible for promoter activity, seed specificity, and temporal regulation of the pea legA gene. Sequences between positions -549 and -1203 bp appear to function as enhancer-like elements, to increase expression.
Shirsat
A
A
Department of Botany, University of Durham, UK.
Wilford
N
N
Croy
R
R
Boulter
D
D
D001483:Base Sequence; D015152:Blotting, Northern; D015139:Blotting, Southern; D002872:Chromosome Deletion; D004797:Enzyme-Linked Immunosorbent Assay; D007887:Fabaceae / Q000235:genetics; D005796:Genes; D007700:Kinetics; D008969:Molecular Sequence Data; D010940:Plant Proteins; D014674:Plant Proteins, Dietary / Q000032:analysis / Q000235:genetics*; D010944:Plants / Q000235:genetics*; D010946:Plants, Medicinal; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D014026:Nicotiana / Q000235:genetics; D000097793:Legumins
D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D000097793:Legumins