基于PCR的非酶连型基因组步移技术研究进展  

Progress on Ligation-independent PCR-mediated Approaches for Genomic Walking

作　　者：李旭娟[1] 陈杨玲[1] 王海波[1,2] 龚明[1] 邹竹荣[1] 

机构地区：[1]云南师范大学生命科学学院,教育部生物质能源持续发展和应用工程研究中心,云南省生物质能源和环境生物技术重点实验室,昆明650500 [2]曲靖师范学院,云南曲靖655011

出　　处：《中国农业科技导报》2013年第2期193-199,共7页Journal of Agricultural Science and Technology

基　　金：国家自然科学基金项目(31060160;31160169);云南省教育厅科研基金项目(2010Z087)资助

摘　　要：基于PCR的基因组步移(或染色体步移)技术常用于克隆已知DNA片段的旁侧序列,是一项重要的的基因组学研究技术。根据原理可将其分为依赖酶连介导和不需酶连介导两大类。目前后者发展迅速,方法种类较多而且各具特色,但根据步移引物与模板DNA的结合特点,可将其统分为基于简并引物的或依赖特定位点的基因组步移技术。综述了当前主要或典型的非酶连PCR介导的基因组步移方法以及最新进展,以期为相关研究提供方法借鉴。PCR-mediated approaches for genomic walking （or chromosome walking） are routinely used for cloning flanking sequences of the known DNA regions, which is an important technology for genomic research. According to the principles, they can be categorized into two types of ligation-dependence and ligation-independence. At present, numerous approaches of the later, each with distinct features, have been rapidly developed. Nevertheless, regarding the binding patterns of walking primers on DNA templates, they can be overall grouped into the degenerate primer- based or specific sequence-dependent subsets. Following this viewpoint, this paper summarized the major or representative techniques and the recent developments of ligation-independent, PCR-mediated genomic walking, so as to provide a methodological reference for the relevant researches.

关 键 词：非酶连 PCR 基因组步移 染色体步移 

分 类 号：Q819[生物学—生物工程]