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

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。

Expression of a Tobacco Glycosyltransferase Gene's Promoter in Transgenic Tobacco

Objective]The aim was to study the expression of the deletion fragments from the promoter of a glycosyltransferase gene induced both by MeJA and SA cloned from Tobacco W38(sm-Ngt) in the trangenic Tobacco.Method] The T1 transgenic Tobacco seedling with the deletion fragments from the promoter of sm-Ngt and Gus gene was treated with MeJA and SA for 16 h,and then histochemical staining and fluorometric analysis of GUS activity of the transgenic plants T1 were carried out to determine GUS activity and analyze the effection of MeJA and SA on expression of the 5 deletion fragments from the promoter of sm-Ngt. Result] In the 5 transgenic T1 Tobacco seedling with the different deletion fragments from the promoter of sm-Ngt,histochemical staining of GUS activity in the transgenic plant with-220-0 bp fragment was least,and with-524-0 and-468-0 bp fragments were the most.In the condition of untreated with MeJA and SA,the GUS activity in the transgenic tobacco seedlings growing 30 days with-524-0 and-468-0 bp were far outclass those with-1 150-0,-800-0 and-220-0 bp,furthermore,this was not because of the copy number.The GUS activity in the transgenic plants with-800-0 bp were induced by both MeJA and SA,and the-1 150-0 bp fragment was induced only by MeJA.Conclusion]There was element exsited that enhancing the expression of sm-Ngtin-524——220 bp of the promoter,and restraining element in-1 150——524 bp region.And in the-1 150-524 bp region there existed many elements induced by both MeJA and SA.