昆虫防御素A的分子改造及其在噬菌体表面的展示  

Rational design of the insect defensin A and its displaying on the surface of phage

作　　者：赵安[1] 薛沿宁[1] 冯健男[1] 张杰[1] 高波[1] 刘农乐[1] 王芳[1] 王会信[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学科学院院刊》2002年第4期270-272,293,共4页Bulletin of the Academy of Military Medical Sciences

基　　金：国家"8 63"计划青年基金课题

摘　　要：目的 :实现对昆虫防御素A的分子改造及改造后分子的噬菌体表面展示 ,为进一步设计构象型肽库 ,研究分子间相互作用奠定基础。方法 :分子改造包括以下内容 :分子截短 ,突变碱性氨基酸 ,减少半胱氨酸的数目等。利用InsightⅡHomology系统对改造后分子结构进行预测。然后再将合成的改造蛋白基因插入到pCANTAB 5E载体中 ,采用Western印迹鉴定该蛋白在噬菌体表面展示。结果与结论 :模建结果显示改造后蛋白仍保持原有三维结构不变 。Objective:To design conformationally constrained peptide library,to realize optimization and display of the insect defensin A on the surface of the phage and to study the interaction of the molecules.Methods:The protein molecule design included truncation at N_terminal, replacement of basic amino acid and reduction of the number of cysteine. InsightⅡ Homology system was used to predict the structure of the changed molecule. The gene of the changed molecule was synthesized and inserted into the pCANTAB 5E vector for phage display. Western blotting was used to detect the expression of insect defensin A on the surface of the phage. Results and Conclusion:The reconstructed molecule retained the parental structure and could be displayed on the surface of the phage successfully.

关 键 词：昆虫防御素A 分子设计 噬菌体展示 

分 类 号：Q819[生物学—生物工程]