一个烟草糖基转移酶启动子在转基因烟草植株中的表达分析Expression of a Tobacco Glycosyltransferase Gene's Promoter in Transgenic Tobacco
王雪;谭艳平;周杰;王春台;刘学群;
摘要(Abstract):
[目的]研究从烟草中克隆的1个受水杨酸和茉莉酸甲酯诱导表达的新糖基转移酶基因(sm-Ngt)启动子部分缺失片段在烟草中的表达。[方法]以转sm-Ngt5个不同长度的5’端缺失的启动子与Gus基因融合植物表达载体的T1代转基因植株为材料,用茉莉酸甲酯(MeJA)和水杨酸(SA)处理16 h后分别进行GUS组织化学染色和荧光定量法测定GUS酶活性,分析水杨酸和茉莉酸甲酯对sm-Ngt5个不同长度的5’端缺失的启动子表达的影响。[结果]在5个不同缺失片段启动子的转基因T1代生长30 d的植株中,转-220~0 bp片段的GUS染色最少,转-524~0 bp及-468~0 bp片段的染色最深。在没有MeJA和SA诱导处理时,-524~0 bp和-468~0 bp 2个启动子片段启动的GUS活性最高,远高于-1 150~0、-800~0、-220~0 bp片段的活性,并且不是由于基因拷贝数而引起的(South-ern杂交结果);-800~0 bp启动子片段启动的GUS活性受到MeJA和SA双重诱导,-1 150~0 bp启动子片段启动的GUS活性受到MeJA的诱导。[结论]在sm-Ngt启动子的-524~-220 bp存在提高启动子活性调控元件,-1 150~-524 bp存在抑制启动子活性的序列,并且存在MeJA和SA双重诱导启动子活性调控元件。
关键词(KeyWords): 烟草;sm-Ngt启动子;茉莉酸甲酯;水杨酸;表达
基金项目(Foundation): 湖北省自然科学基金(2004ABA123)
作者(Author): 王雪;谭艳平;周杰;王春台;刘学群;
Email:
DOI: 10.13989/j.cnki.0517-6611.2010.18.092
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