噬菌体展示技术与其应用.pptVIP

示技术及其应用 食品学院廖振林 Email: liaoOshenli caml  噬菌体展示技术是将 各种多肽或蛋白质以融合 蛋白的形式表达并展示在 噬菌体表面,同时将其遗 传密码包含于噬菌体内部 这使得蛋白质的功能与其 基因密码有机的连结在 起 2014-3-30  非展示系统 展示系统 CCsc 2014-3-30  The expression of exogenous peptides on the surface of filamentous bacteriophage was first study, different molecules such as sma/ o initially described by Smith in 1985. Since hi peptides and antibodies have been displayed on coat proteins of phage, greatly expandin the applications of the technology. The past decade has seen considerable progress in the techniques and applications of phage libraries In addition different screening methods have allowed isolation and characterization of peptides binding to several molecules in vitro in the context of living cells, in animals and in humans 2014-3-30  In vivo use of phage libraries. Initially the phage library is injected in the circulation Next, phage is allowed to circulate for Tail vein injection minutes or hours (in this case when of phage internalized phages are to be recovered) Finally, after deep anesthesia euthanized and the organs are cted phage recovery and peptide sequencing Phage recovery In vivo binding Peptide sequencing  1+ Phage structure. PIll pVI, pvIl, pvIl and pXIX represent phage proteins Exogenous peptides are expressed or display usually on pIll or pill 2014-3-30  丝状噬菌体的生物学 ()p{5 p5)X(b) (d)schematic representation af M13(not to scale) 形态长杆状,长度约1um,管直径约6mm 基因组:约640核苷酸大小的单链线状DNA 结构:●2700个主要蛋白(Pvm)分子螺旋状排列形成长管, 头5个拷贝的次要壳蛋白PI和PVI, ●另一头则由次要壳蛋白PVI和PIX。 生理:●F菌毛结合的序列是PI蛋白N端的200个氨基酸 个分裂周期中可生产几百个子代 3其产量可达到0.3mg/m  源蛋白的展示部位 hM· 已m】 Mild type phage Phago die Phage displying 图2外源發白在噬菌体登白中的融合位置 2014-3-30  噬菌体展示技术所展示外源多肽和及文库 随机肽库 肽库通常较短(4-36氨基酸9长度) 它们的核酸由化学法合成而得 示方式包括3,33,3+3,8,88,8+8等 因文库 基抗病一 Natural Synthetic peptides random peptides 3.各种自然蛋白及蛋白片段等 素如蓖麻 物质、神 结合蛋白如锌指蛋 白等;酶底物如蛋白酶底物等;细胞素如IL3,IL6等。 2014-3-30  抗体库技术简单流程 2014-3-30