基于荧光激活细胞分选技术的超高通量酶活性筛选方法及其应用  

Ultrahigh-throughput Enzymatic Screening Method Based on Fluorescence-activated Cell Sorting and Its Applications

作　　者：马富强[1,2] 冯雁[1,2] 杨广宇[1] 

机构地区：[1]上海交通大学生命科学技术学院,上海200240 [2]吉林大学生命科学学院,长春130012

出　　处：《生物化学与生物物理进展》2012年第4期299-306,共8页Progress In Biochemistry and Biophysics

基　　金：国家重点基础研究发展计划(973)(2011CBA00800);国家自然科学基金(30970632;31070056;30821005)资助项目~~

摘　　要：基于荧光激活细胞分选(FACS)技术的超高通量酶活性筛选方法是新出现的一类高通量筛选技术.它利用流式细胞仪高灵敏度、高通量的特点,能以极高的速度(>108/天)对大容量酶基因文库进行筛选.FACS筛选技术的出现突破了常规筛选方法低效、耗时、费力等瓶颈问题,极大地提升了人类对大容量基因文库的探索能力,因此在新酶基因筛选、酶活性检测、酶定向进化等领域有广泛的应用潜力.综述了FACS超高通量酶活性筛选方法的最新研究进展,着重介绍了其在酶定向进化中的应用.Due to its super sensitivity and incomparable high throughput,fluorescence-activated cell sorting（FACS） based screening method is emerging as an important ultrahigh-throughput enzymatic screening technology recently.It could detect multiple fluorescent parameters simultaneously and screen enzyme libraries at an extremely high speed（ 〉10^8/d）,thus possesses many advantages over the conventional technologies.FACS based screening method greatly improves our ability in handling large protein libraries and has tremendous potential in various fields such as metagenomics and directed evolution.Herein,recent progress of FACS based enzymatic screening method has been reviewed,with an emphasis on its applications in enzyme directed evolution.

关 键 词：荧光激活细胞分选 高通量筛选 酶 定向进化 

分 类 号：Q503[生物学—生物化学] Q789