Xie Yingqiu, Liu Yule, Meng Meng, Chen Lei, Zhu Zhen
Laboratory of Plant Genetic Manipulation, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China.
Plant Mol Biol. 2003 Sep;53(1-2):1-14. doi: 10.1023/B:PLAN.0000009257.37471.02.
The activity of the C1 and the V1 gene promoter of cotton leaf curl Multan virus (CLCuMV) was investigated in transgenic plants with the gus gene as a reporter gene. Quantitative GUS activity analysis of the transgenic plant leaves showed the average activity of the CLCuMV C1 gene promoter was 3- to 5-fold higher than that of the CaMV 35S promoter, with maximal expression being 10-fold higher. CLCuMV V1 gene promoter activity was only about 1/10th that of the CaMV 35S promoter in the absence of trans-activator C2. Histochemical GUS staining of the transgenic plants indicated that the CLCuMV C1 gene promoter was active in leaves, stems, roots and almost all reproductive organs. Functional analysis of promoter 5'-deletion series indicated that promoter activity of a 257 nucleotide fragment (-257 to the transcription initiation site) and a 241 nucleotide fragment (-241 to the transcription initiation site) were 5-fold and 2-fold stronger than that of the full-length CLCuMV C1 promoter respectively. These results demonstrate that the CLCuMV C1 promoter is a super-strong near-constitutive promoter in plants and has great application potential for plant genetic engineering studies.
以gus基因作为报告基因，在转基因植物中研究了棉花曲叶木尔坦病毒（CLCuMV）的C1和V1基因启动子的活性。对转基因植物叶片进行的定量GUS活性分析表明，CLCuMV C1基因启动子的平均活性比花椰菜花叶病毒（CaMV）35S启动子高3至5倍，最大表达量高10倍。在没有反式激活因子C2的情况下，CLCuMV V1基因启动子活性仅约为CaMV 35S启动子的1/10。转基因植物的组织化学GUS染色表明，CLCuMV C1基因启动子在叶、茎、根以及几乎所有生殖器官中均有活性。启动子5'缺失系列的功能分析表明，一个257个核苷酸片段（-257至转录起始位点）和一个241个核苷酸片段（-241至转录起始位点）的启动子活性分别比全长CLCuMV C1启动子强5倍和2倍。这些结果表明，CLCuMV C1启动子是植物中一种超强的近组成型启动子，在植物基因工程研究中具有巨大的应用潜力。
