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目的分析研究黄花蒿3-羟基-3-甲基戊二酰辅酶A还原酶(HMGR)启动子的活性表达及其核心序列。方法以β-葡萄糖苷酸酶(GUS)基因作为报告基因,通过构建包括HMGR启动子全长序列在内的5个不同长度表达载体,并采用农杆菌介导法转化烟草以及对转化烟草进行GUS染色。结果转HMGR启动子烟草的根、茎、叶、花、果荚均发现蓝色;脱水和高温胁迫会降低该启动子的表达,低温胁迫对其影响不明显;启动子片段Aa GPX1、Aa GPX2和Aa GPX3具有驱动GUS报告基因表达的功能,而Aa GPX4和Aa GPX5则未有此功能。结论 HMGR启动子在烟草整个生长期内均能够稳定表达;该启动子对低温不敏感,而对高温和脱水条件敏感;该启动子的核心序列在P-HMGR-3区域,而Aa GPX2与Aa GPX3片段的非重叠区则存在与HMGR启动子活性强弱相关的调控元件。
OBJECTIVE: To study the active expression and its core sequence of 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) promoter of Artemisia annua. Methods The GUS gene was used as reporter gene. Five different length expression vectors including the full-length HMGR promoter were constructed. Agrobacterium-mediated transformation of tobacco and GUS dyeing. Results The HMGR promoter was found to be blue in roots, stems, leaves, flowers and pods. Dehydration and high temperature stress reduced the expression of the promoter, but the effect was not obvious under low temperature stress. The promoter fragments Aa GPX1, Aa GPX2 And Aa GPX3 function to drive GUS reporter expression, whereas Aa GPX4 and Aa GPX5 do not. Conclusion HMGR promoter can be stably expressed in tobacco throughout the growth period; the promoter is insensitive to low temperature and sensitive to high temperature and dehydration conditions; the promoter sequence of the promoter is in the P-HMGR-3 region, whereas Aa GPX2 and Aa The non-overlapping region of GPX3 has regulatory elements related to the activity of HMGR promoter.