利用EvolvR系统对酶基因实现多窗口定向进化的研究  

Research on the multi-windows directed evolution of target enzyme gene using EvolvR system

作　　者：林峻[1] 陈珏 黄佳敏 LIN Jun;CHEN Jue;HUANG Jiamin(Institute of Applied Genomics,College of Biological Science and Engineering,Fuzhou University,Fuzhou,Fujian 350108,China)

机构地区：[1]福州大学生物科学与工程学院,福州大学应用基因组学研究所,福建福州350108

出　　处：《福州大学学报（自然科学版）》2021年第2期270-276,共7页Journal of Fuzhou University(Natural Science Edition)

基　　金：福建省科技厅引导性项目(2020Y0003);福建省卫生计生委青年科研课题(2017-1-62);福建省科技创新联合项目(2018Y9105);福州大学贵重仪器设备开放测试基金资助项目(2020T020)。

摘　　要：EvolvR系统是一种CRISPR介导的新型定向进化技术,本研究探索其在酶基因序列定向进化中的适用性以及突变效率,同时在此基础上扩大EvolvR的突变窗口长度.研究成功将4个能高效表达的单个sgRNA串联,并检测到靶向同一基因的3个不同靶位点.证明EvolvR具有在目标基因区域大范围制造突变的潜力,具有很高的应用价值.为利用EvolvR系统对目标酶基因实现定向进化的研究奠定基础.EvolvR is a new CRISPR-mediated directed evolution technology.This study aimed to explore the applicability and mutation efficiency of the EvolvR system in the directed evolution of enzymes and the length of the mutation window.The results showed that concatenation of four efficiently expressed individual sgRNA could indeed target three sites of a same gene.EvolvR was proved to have the potential to produce mutations in a wide range of target gene regions and had high application value.This study laid a foundation for the targeted enzyme directed evolution with EvolvR system.

关 键 词：定向进化 EvolvR 随机突变 突变文库 

分 类 号：Q518.4[生物学—生物化学]