虫草多糖荧光标记的方法学研究

Fluorescent Substance Labeling of Cordyceps Polysaccharide

目的:对人工培养虫草多糖进行荧光标记,以供多糖的有关生物学研究。方法:先将多糖与酪胺反应,然后再与异硫氰酸荧光素(FITC)反应。对人工培养虫草多糖与酪胺反应中的反应时间、反应温度、pH、和酪胺用量进行了单因素考察。280nm的吸收峰表示多糖与酪胺的结合;450nm的吸收峰表示与FITC的结合。结果与结论:多糖荧光标记的最佳条件是:反应时间越长,温度在60℃,pH为8时反应率最高。其他多糖的荧光标记得出结构复杂的多糖,反应效率要低。

Objective：Fluorescent labeling of polysaccharides from artificial cultured cordycepin was developed for further bioresearch of polysaccharide.Methods：The end of the CP was reacted with tyramine（Tyr） to synthesize CP-Tyr,then the compound was reacted with fluorescein isothiocyanate（FITC）.We studied the effects of the reaction time,temperature,PH,the amount of tyramine on CP-Tyr formation.Results and Conclusion：CP itself had no absorption at 280nm,CP-Tyr had absorption at 280nm,CP-tyramine-FITC had absorption at 280nm and 450nm.Single-factor studies showed that reaction time had the greatest impact,the other factors had no significant effects.of the,The reaction efficiency was low for polysaccharides with complex structures.The optimal condition for fluorescent labeling of cordycepin was 60℃ and pH8.0.The UV absorption proved that fluorescent material had attached to Cordyceps polysaccharides.Comparison of fluorescent labeling between different polysaccharides suggested that the more complicate of structure lead to the lower reaction efficiency.