摘 要 ：为确定拟南芥抗逆相关基因AtRPK1启动子的顺式功能元件，对其启动子区进行了分段克隆。通过5′端缺失方法得到203、316、604、809 bp 4个启动子片段，分别构建成p1300-pro-GUS表达载体，并转入拟南芥，进行GUS染色和GUS定量检测。通过对809 bp全长启动子转基因拟南芥GUS染色发现，转基因拟南芥的叶片、茎、花、根中均有表达，在分生能力强的组织和维管束集中的组织，AtRPK1基因启动子具有较高启动表达能力。5′端缺失启动子检测结果表明，转录起始点到启动子上游114位点区域包含AtRPK1基因启动子的关键顺式作用元件。对启动子缺失片段转基因植株利用200 mmol·L-1 NaCl胁迫3 h后，β-葡萄糖苷酸酶活力定量检测结果表明，在启动子上游-19位点处的GT-1顺式作用元件GAAAAA可能直接与盐胁迫应答相关。
Abstract:To identify the critical cis-acting element, the deletion analysis was performed using the promoter region of the stress resistance related gene AtRPK1 in Arabidopsis. By the 5′ terminal deletion methods, the 203, 316, 604, 809 bp promoter fragments were obtained and constructed into p1300-pro-GUS expression vector and transferred into Arabidopsis. By GUS staining detection of 809 bp length promoter transgenic Arabidopsis, it was found that the AtRPK1 gene was expressed in the leaves, stems, flowers and roots. However, the expression of AtRPK1 was stronger in meristem organization and vascular tissue. The detection results show that the region from the transcription start sites to the -114 section of AtRPK1 gene promoter contains the critical cisacting elements. After the stress of 200 mmol·L-1 NaCl, quantification detection results of β-glucuronidase activity showed that the GT-1 cis-element GAAAAA at -19 section may be directly related to the salt stress response.