番木瓜诱导型基因CpPGIP2启动子的克隆与分析Isolation and analysis of the promoter for an inducible gene CpPGIP2 from Carica Papaya
申艳红;陈晓静;戴志林;
摘要(Abstract):
【目的】获得诱导型基因Cp PGIP2启动子,并分析该启动子的功能。【方法】以番木瓜Cp PGIP2基因c DNA序列(HQ660394)搜索番木瓜基因组DNA序列,获得一段约2 000 bp的5’端上游DNA序列(ABIM01005077),参照该序列设计PCR特异引物,以番木瓜叶片DNA为模板克隆该启动子,并进行生物信息学分析。将3个启动子片段替换p CAMBIA1301载体中的Ca MV 35S启动子,构建缺失启动子表达载体,以GUS瞬时表达检测缺失启动子表达活性。【结果】获得了Cp PGIP2起始密码子上游长为1 981 bp的调控序列,经分析该序列含有真菌、脱落酸、赤霉素、光照等多种响应元件,为诱导型启动子。构建了1个启动子全长表达载体和2个5’端缺失启动子表达载体,分别命名为p D0-1981、p D1-1204和p D2-261。启动子在番木瓜果肉中的瞬时表达显示,长度为1 204 bp的启动子表达活性最强。并且,该启动子在根、茎、叶、果肉和愈伤组织中均有不同程度的表达,在近外果皮的果肉处和根部表达最明显。【结论】本研究获得了Cp PGIP2启动子序列,确定了表达活性最强的启动子长度,初步分析了启动子的表达模式和顺式作用元件,为进一步深入研究Cp PGIP2基因功能以及将该启动子应用于植物基因工程育种奠定了基础。
关键词(KeyWords): 番木瓜;多聚半乳糖醛酸酶抑制蛋白;启动子;调控元件;载体构建
基金项目(Foundation): 福建农林大学园艺学院青年学术骨干培养基金(FAFU2012YYPY05);; 福建省自然科学基金资助项目(2010J05048)
作者(Author): 申艳红;陈晓静;戴志林;
Email:
DOI: 10.13925/j.cnki.gsxb.20150180
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