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作 者 ：叶冰莹;王婷;何文锦;邱思;陈由强;*

期 刊 ：植物研究 2014年 34卷 1期 页码：62-67

关键词：5侧翼序列；缺失表达；甘蔗；UGPase；

Keywords:5′ flanking sequences, loss expression, Sugarcane, UGPase,

摘 要 ：以甘蔗(FN95;-1702)为材料，通过接头连接PCR方法克隆该基因不同长度5′侧翼序列。将不同长度的5′侧翼序列连同UGPase基因的外显子片段定向插入到GUS基因上游，在保证其后GUS编码框不发生偏移的情况下，插入的UGPase外显子融合GUS表达成为新的报告基因。根据此策略，构建了一系列表达结构为5′ Flanking Sequence-UGPase Exon-GUS-Nos polyA的5′侧翼序列缺失表达载体，进行启动子活性分析。注射法转染烟草叶片组织检测GUS瞬时表达，分析结果表明，所克隆到的UGPase基因5′端侧翼序列不具有启动子活性。

Abstract:The different lengths of 5′ flanking sequence of UGPase were cloned by Adaptor-ligation PCR. These different lengths of 5′ flanking sequences of UGPase gene were fused with the coding sequence of GUS (β-glucuronidase) gene to construct fusion genes. All the fusion genes were injected into leaves of Nicotiana tabacum for transient GUS expression. The results showed that the 5′ flanking sequence of UGPase gene did not have any promoter activity.

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