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百合花特异启动子PchsA表达载体的构建及功能分析
常小丽1, 刘雅莉1, 王跃进1
西北农林科技大学 园艺学院,农业部西北园艺植物种质资源与遗传改良重点开放实验室
摘要:
[目的]分析从百合中克隆获得的查耳酮合成酶基因(chsA)启动子的组织表达特性.[方法]将chsA启动子片段插入到双元表达载体pCAMBIA 1381中,成功构建了植物表达载体pCAMBIA-CHS,通过根癌农杆菌EHA 105转化拟南芥,用50 mg/L潮霉素对转化植株进行筛选,对阳性植株进行PCR检测和GUS组织化学分析.[结果]PCR检测表明,chsA启动子片段已经整合到拟南芥基因组中,GUS组织化学分析显示,在拟南芥花序中有很强的GUS活性,叶腋处有微量表达,其他组织中不表达.[结论]初步证明百合chsA启动子为花特异表达启动子.
关键词:  启动子  GUS基因  植物表达载体  拟南芥转化
DOI:
分类号:
基金项目:国家自然科学基金?
Construction and function analysis of the expression vector of the flower-specific promoter PchsA from lily
Abstract:
【Objective】 This paper aimed to verify the tissue specific expression of chsA promoter cloned from Lily.【Method】 The chsA promoter was constructed into the binary vector pCAMBIA 1381 and introduced into Arabidopsis thaliana by Agrobacterium mediated method.The transformed plants were selected by 50 mg/L HygromycinB stress.Analysis of PCR assay and GUS activities were employed to screen the specific expression.【Result】 PCR analysis of the positive plants showed that the promoter had been integrated into the Arabidopsis genome.The determinations of the GUS activities indicated that the chsA promoter could drive the GUS reporter gene to express in the flower of transgenic plants,while no or very weak expression of the GUS reporter gene in other tissues.【Conclusion】 The chsA promoter is supposed to be related to flower-specific expression.
Key words:  chsA promoter  GUS gene  plant expression vector  Arabidopsis thaliana transformation