Isolation and characterization of a chitinase gene VCH3 promoter from grapevine (Vitis amurensis)

The 1216-bp 5' upstream region of the gene encoding a class III chitinase VCH3 was isolated from grapevine (Vitis amurensis Rupr.) by adaptor-PCR (GenBank accession number AF441123), and the transcriptional start site of the VCH3 gene was identified by primer extension, which corresponds to the second A in the DNA sequence 5'-ATCAAGCAC-3'. Sequence analysis revealed that the VCH3 promoter sequence contains CAAT and TATA motifs that are located at the -122 and -29 nucleotide upstream of the transcriptional start site, respectively. Both motifs are characteristic of the eukaryotic gene promoter. In addition, within the VCH3 promoter, we found two inverse salicylic acid (SA)-responsive cis-acting motifs (TGACG) at -1181 bp and -293 bp upstream of the transcriptional start site, respectively. To characterize the VCH3 promoter, the VCH3 promoter-GUS chimera was constructed and transferred to Nicotiana tobacum cv. NC89 by Agrobacterium tumefaciens-mediated leaf disc transformation. Fluorometric and histochemical analysis of GUS activity in the transgenic tobacco root and leaf treated with SA showed that the VCH3 promoter was induced by SA, which indicates that it may have potential use in genetic engineering.