CAJ | 학술논문

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.
기우PCR적기인조보이(혹염색체보이)기술상용우극륭이지DNA편단적방측서렬,시일항중요적적기인조학연구기술.근거원리가장기분위의뢰매련개도화불수매련개도량대류.목전후자발전신속,방법충류교다이차각구특색,단근거보이인물여모판DNA적결합특점,가장기통분위기우간병인물적혹의뢰특정위점적기인조보이기술.종술료당전주요혹전형적비매련PCR개도적기인조보이방법이급최신진전,이기위상관연구제공방법차감.
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.