Flanking genomic sequences refer to the DNA sequences flanking specific sites of known sequences in chromosome, which contain information such as candidate genes, transcriptional regulation, chromosome structure, and biosafety, and play an important role in genomics research. Flanking sequence acquisition technologies are mainly used in the cloning of regulatory sequences such as promoters and enhancers, identification of T-DNA or transposon insertion sites, chromosome walking, genome-wide gap filling, etc. It is an important means of structural genomics research and functional genomics research. It is applied in the identification of transgenic plants and animals and their safety management. With the development of molecular biology, many methods for obtaining flanking sequences have been established, including plasmid rescue, inverse PCR, ligation-mediated PCR, semi-random primer PCR, whole-genome resequencing etc. In this review, we summarize and compared different methods for acquiring flanking genomic sequence. The principles and research progress of each approach are discussed.
侧翼序列是指染色体中特定位点两侧的DNA序列,包含着候选基因、转录调控、染色体结构、生物安全等信息,在基因组学研究中具有重要的作用。侧翼序列获取技术主要应用于启动子和增强子等调控序列的克隆、鉴定T-DNA或转座子插入位点、染色体步移、全基因组空隙填补等,是结构基因组研究以及功能基因组研究的重要手段,在转基因动植物鉴定及安全管理等方面具有重要应用。随着分子生物学的发展,目前已经建立了许多侧翼序列的获取方法,依据技术原理可以分为质粒拯救法、反向PCR法、外源接头介导PCR法、半随机引物PCR法和基因组重测序法等5大类。本文系统总结了近年来侧翼序列获取技术的研究进展,并对这些技术的原理以及应用情况进行了较为系统的综述,为侧翼序列信息的获取提供参考。.
Keywords: flanking sequence; inverse PCR; ligation-mediated PCR; plasmid rescue; semi-random primer PCR; whole-genome resequencing.