中国生物工程杂志

Based on the previously reported sequence of soybean 1-aminocyclopr- opane-1-carboxylate synthase gene (GenBank Accession No.dq273840), three gene-specific reverse primers, CTA798 sp1, CTA798 sp-2, CTA798 sp3 were designed, and a DNA fragment about 600bp was obtained by TAIL-PCR. The transcription start site was localized at 62bp upstream of the start ATG identified by 5′-RACE. The obtained sequence was analyzed by PLACE and PLANTCARE program and showed putative ACC synthase gene promoter features: it contains the basic core elements of TATA-box and CAAT-box, as well as stress-induced elements: light-responsive element Box1, Box4 and elicitor-induced element W-box. Based on the binary vector pBI121, a plant gene expression vector carrying gusA gene under the putative ACC synthase gene promoter was constructed and introduced into Agrobacterium tumefaciens EH105. The transient expression of gusA gene was then observed by histochemical staining in tobacco leaves through the leaf disc transformation mediated by Agrobacterium tumefaciens, which revealed the obtained sequence has promoter activity.