Gateway克隆技术背景原理与其应用.pdf

Gateway® 克隆技术背景、 原理及其应用 王宇、余辉 首都师范大学 Table of Contents 1 Overview 2 Advahtages of the Gateway Technology 3 Application of Gateway Technology 4 Examples What is “Gateway Technology”? ‘The Gateway® Technology is a universal cloning method based on the site-specific recombination properties of bacteriophage lambda.’ Gateway® Technology Handbook . Invtrogen® Keyword：universal、site-specific recombination、 bacteriophage lambda Genetic Recombination Defination of Genetic Recombination: ‘Genetic recombination is a process by which a molecule of nucleic acid (usually DNA, but can also be RNA) is broken and then joined to a different one.’ en.wikipedia.org Types of Genetic Recombination： Homologous recombination （同源重组） Site-specific recombination （位点专一性重组） Homologous Recombination （同源重组）： ‘Recombination that requires extensive sequence similarity between the recombining DNAs.’ Robert F.Weaver .Molecular Biology 3th ‘同源重组是更为普遍的一种重组机制，它可以发生在任何两种具有相 同或相似序列之间，涉及到两个DNA分子在相同区域的断裂和重新连 接。’ Characteristic of Homologous Recombination Random site （发生在随机位点） Require extensive sequence similarity （需要一定长度的 同源片段，1kb） Site-specific recombination （位点专一性重组）: ‘Recombination that always occurs in the same place and depends on limited sequence similarity between the recombining DNAs.’ Robert F.Weaver .Molecular Biology 3th ‘重组反应发生在两个特异序列（不一定同源）之间，如噬菌体整合/ 外切或转座中共合体结构的解离。’ Benjamin Lewin .Genes VIII Bacteriophage lambda （λ噬菌体） 噬菌体生活史 ： 溶原状态 →裂解状态（整合integration） 裂解状态 →溶原状态（外切excision） 整合（Int、IHF） 溶原状态