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  • 1
    ISSN: 1617-4623
    Keywords: Chlorophyll a/b binding protein genes ; Divergent promoters ; Coordinate expression ; Between-transformant variability
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We examined the divergent Cab22R and Cab22L genes of petunia as a coupled photosynthetic promoter casstte for simultaneously directing high level expression of two foreign genes in a coordinated and tissue specific manner in transgenic plants. Primer extension assays with gene specific primers on petunia leaf RNA indicated that abundant and coordinated mRNA expression occurs for the two Cab22 divergent transcription units in a tissue specific and light regulated fashion. The expression characteristics of the intact Cab22 genes can be maintained in transgenic tobacco plants, but are influenced by the site of insertion into the Agrobacterium T-DNA binary vector. Thus, the expression levels of genes introduced near the T-DNA left border varied coordinatedly among independently transformed plants and remained completely light inducible and tissue specific. In contrast, each of these transcription properties was significantly disrupted by the insertion of the genes into the right section of the binary vector, between a selectable NPTII gene and the T-DNA octopine synthase (Ocs) cis-acting promoter element. Gene transfer experiments with chimeric gene constructs have shown that both the quantitative and qualitative expression properties of the Cab22 genes can be conferred to foreign protein coding genes via fusion to the Cab22 leader and upstream sequences. The use of this divergent promoter as a cassette to express foreign genes in plants in a coordinated fashion and the possible influences of differential mRNA stabilities and transcription rates between intact and chimeric genes in transgenic plants are discussed.
    Type of Medium: Electronic Resource
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  • 2
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 215 (1989), S. 337-344 
    ISSN: 1617-4623
    Keywords: Divergent promoters ; Chlorophyll a/b binding protein genes ; cis-acting elements ; RNA expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary In order to identify specific cis-acting elements which regulate the expression of the divergent Cab22R and Cab22L genes of Petunia, we conducted systematic mutational studies of the 1 kb intergenic promoter region. Sequence analysis revealed three GATA box sequence repeats positioned between the TATA and CAAT box elements. These GATA elements are conserved in corresponding promoter regions of all LHCII Type I Cab genes in Petunia and other dicotyledonous plants we have examined. Sitespecific mutations in the CAAT box and the GATA box elements of the Cab22R promoter resulted in 8-fold and 5-fold reductions in Cab22R transcript levels respectively. A deletion of 52 bp, adjacent and upstream from the CAAT box (-92 to- 145) in the Cab22R promoter reduced transcript levels 20-fold. This deletion contains a region of 13 pb which is conserved between many Petunia Cab genes. These results indicate that the quantitative expression of the Cab22 promoters is regulated by multiple cis-acting elements including CAAT and GATA box elements as well as sequences located between -92 and -145. The deletion of the region between -92 and -145 is partially compensted by homologous sequences present in the adjacent divergent promoter Cab22L.
    Type of Medium: Electronic Resource
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