Abstract
Using a modified TAIL-PCR technique, the 5′-flanking regions of the phenylalanine ammonia lyase (Pal) genes of a yam species, Dioscorea bulbifera, and the phosphoglucose isomerase (Pgi) gene of D. tokoro were successfully isolated. Two novel modifications of the TAIL-PCR procedure introduced here, namely (1) the use of a battery of random 10-mers (RAPD primers) as short arbitrary primers, and (2) the use of a total of five nested, gene-specific primers, allow the rapid isolation of the 5′-flanking region of any gene from organisms with large genomes. Isolated 5′-flanking regions were fused to the gus gene, and tested for transient expression in tobacco BY2 cells. All the isolated 5′-flanking regions were shown to drive reporter gene expression. Three Pal promoters responded to salicylic acid, presumably as a result of the binding of a MYB transcriptional activator to the multiple MREs (Myb Recognition Elements) present in these regions.
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Received: 10 October 1999 / Accepted: 31 January 2000
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Terauchi, R., Kahl, G. Rapid isolation of promoter sequences by TAIL-PCR: the 5′-flanking regions of Pal and Pgi genes from yams (Dioscorea). Mol Gen Genet 263, 554–560 (2000). https://doi.org/10.1007/s004380051201
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DOI: https://doi.org/10.1007/s004380051201