楸子S6PDH基因启动子活性鉴定

利用同源PCR技术从楸子(Malus prunifolia)基因组DNA中扩增得到6-磷酸山梨醇脱氢酶(Sorbitol-6-phosphate dehydrogenase,S6PDH)基因的启动子并命名为Mp-S-P。对启动子序列的顺式作用元件进行预测分析发现,Mp-S-P序列中除了含有植物启动子的基本结构元件外,还有多个与胁迫和激素诱导相关的元件。以β-葡萄糖苷酸酶(beta-glucuronidase,GUS)作为报告基因,构建含有pMp-S-P-GUS的融合表达载体并通过根癌农杆菌瞬时转化烟草叶片,然后分析该启动子对不同外界条件的响应。结果表明:低温、水杨酸和赤霉素处理可以提高该启动子的活性,而避光、脱落酸和茉莉酸甲酯可以降低该启动子的活性。

Activity Analysis of S6PDH Promoter from Malus prunifolia

The promoter of Sorbitol-6-phosphate dehydrogenase was amplified from genomic DNA of Malus prunifolia,named Mp-S-P. The bioinformatic analysis of promoter sequence was carried out based on plant cis-acting element database. The results indicated that expect for the conservative structural elements,Mp-S-P also contained some defense-related and hormone-induced cis-elements. A fusion expression vector contained Mp-S-P and beta-glucuronidase (GUS) were transiently transformed in leaves of tobacco by agrobacterium GV3101 to analyze the response of Mp-S-P against different treatments. The results revealed that the Mp-S-P activity JP2]could be improved by low temperature,salicylic acid and gibberellin,while weakened by shading,abscisic acid and jasmonic acid methyl ester.