Strong negative and positive regulatory elements contribute to the high-level fruit-specific expression of the tomato 2A11 gene.
Fruit-specific expression of beta-glucuronidase (GUS) activity was produced in transgenic tomato plants when the GUS-coding region was flanked by 5' and 3' regions of the tomato 2A11 gene. Deletion studies on the 5' region revealed a number of strong regulatory elements involved in the proper expression of the 2A11 gene. A 4.0 kb and a 1.3 kb 5' region can confer high-level fruit-specific GUS expression, while a 1.8 kb 5' region produces no GUS activity in leaf or fruit tissue. Thus, a strong negative regulatory element is present in the region between 1324 bp and 1796 bp upstream of the 2A11 transcriptional start and a strong fruit-specific positive regulatory element is present more than 1.8 kb upstream of the transcriptional start site. The 1.8 kb promoter region can be activated by the upstream insertion of the CaMV 35S enhancer sequence, albeit not in a fruit-specific fashion. Substitution of the 3' region of the 2A11 gene with a different 3' region does not seem to affect GUS expression significantly, indicating a minor role, if any, for the 3' region in the fruit-specific expression of the 2A11 gene.
Van Haaren
M J
MJ
Calgene Inc., Davis, CA 95616.
Houck
C M
CM
D016960:Agrobacterium tumefaciens / Q000235:genetics; D001483:Base Sequence; D015152:Blotting, Northern; D004252:DNA Mutational Analysis; D004251:DNA Transposable Elements / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D004742:Enhancer Elements, Genetic / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics; D005638:Fruit / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D015870:Gene Expression / Q000502:physiology; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D007612:Kanamycin / Q000494:pharmacology; D008969:Molecular Sequence Data; D009029:Mosaic Viruses / Q000235:genetics; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000201:enzymology / Q000235:genetics / Q000254:growth & development; D011401:Promoter Regions, Genetic / Q000235:genetics; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D015183:Restriction Mapping
D004251:DNA Transposable Elements; D010940:Plant Proteins; D007612:Kanamycin; D005966:Glucuronidase