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为了寻找天然来源的光敏剂,通过色谱技术,从五叶山小桔中分离出4个生物碱,它们的结构通过波谱数据分别鉴定为glycoborinine(1)、glybomine B(2)、carbalexin A(3)和N-p-coumaroyltyramine(4)。通过薄层色谱自显影技术,评价了4个生物碱对金葡菌和枯草芽孢杆菌的光活化抗微生物的活性,化合物1和4显示出一定的活性。以质粒pET-28为模板,通过PCR反应获得1.8 kb的DNA片段,通过凝胶电泳的方法,研究了4个生物碱类化合物与DNA的结合性,研究表明,化合物1经UVA照射处理能够插入到含有5’-TpA和5’-ApT的DNA序列中,从而对Nde Ⅰ、Nco Ⅰ、Xba Ⅰ和Bcl Ⅰ四种酶对其识别位点的切割显示出一定的抑制活性,与8-甲氧基补骨脂素显示出同样的抑制模式。然而,其他3个化合物对任何限制性内切酶都没有表现出抑制活性。
In order to search for photosensitizers of natural origin, four alkaloids were isolated from Chieh-xiaoyu by chromatographic techniques and their structures were identified as glycoborinine (1), glybomine B (2), carbalexin A (3) And Np-coumaroyltyramine (4). The photoactivatable antimicrobial activity of four alkaloids against Staphylococcus aureus and Bacillus subtilis was evaluated by thin layer chromatography self-imaging technique. Compounds 1 and 4 showed some activity. Using plasmid pET-28 as a template, a 1.8 kb DNA fragment was obtained by PCR and the binding of four alkaloids to DNA was studied by gel electrophoresis. The results showed that Compound 1 could be inserted into the DNA by UVA irradiation To the DNA sequence containing 5’-TpA and 5’-ApT, thus showing some inhibitory activity on the recognition sites of Nde Ⅰ, Nco Ⅰ, Xba Ⅰ and Bcl Ⅰ, Oxy psoralen showed the same mode of inhibition. However, none of the other three compounds showed inhibitory activity against any of the restriction enzymes.