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本研究以获得的丹参SmPAP1基因c DNA为模板设计引物扩增其基因组DNA序列(genomic DNA,g DNA)。g DNA与c DNA序列比对结果显示,该基因基因组DNA序列长度为739 bp,由2个外显子和1个内含子组成。进一步采用染色体步移技术克隆了其上游的一段长度为1 197 bp的启动子序列。启动子序列分析结果显示,该区域内不仅含有保守的TATA盒和CAAT盒,而且包含多个潜在的与胁迫应答相关的顺式作用元件,暗示SmPAP1基因很可能响应多种环境胁迫诱导表达。此外,SmPAP1基因的Southern杂交结果表明,在丹参基因组中SmPAP1仅有1个拷贝数。本研究结果为进一步研究SmPAP1基因的表达调控模式以及启动子的功能奠定基础。
In this study, genomic DNA (gDNA) was amplified by using the c DNA of Salvia miltiorrhiza SmPAP1 gene as template. The results of g DNA and c DNA sequence comparison showed that the length of the genomic DNA was 739 bp, which consisted of two exons and one intron. A further 1 197 bp promoter sequence was cloned by chromosome walking technique. The results of promoter sequence analysis showed that this region contains not only the conserved TATA box and CAAT box, but also a number of potential cis-acting elements involved in stress response, suggesting that SmPAP1 gene is likely to be responsive to various environmental stress-induced expressions. In addition, Southern hybridization results of SmPAP1 gene showed that there was only one copy of SmPAP1 in Salvia miltiorrhiza genome. The results of this study lay the foundation for further study on the expression regulation mode of SmPAP1 gene and the function of promoter.