Rapid isolation of promoter sequences by TAIL-PCR: the 5'-flanking regions of Pal and Pgi genes from yams (Dioscorea)

Mol Gen Genet. 2000 Apr;263(3):554-60. doi: 10.1007/s004380051201.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Base Sequence
  • Exons
  • Glucose-6-Phosphate Isomerase / genetics*
  • Introns
  • Liliaceae / genetics*
  • Models, Genetic
  • Molecular Sequence Data
  • Phenylalanine Ammonia-Lyase / genetics*
  • Polymerase Chain Reaction / methods*
  • Promoter Regions, Genetic*
  • Sequence Analysis, DNA / methods*
  • Sequence Homology, Nucleic Acid


  • Phenylalanine Ammonia-Lyase
  • Glucose-6-Phosphate Isomerase