单分子实时测序技术在环境微生物研究中的应用

Application of single molecule real time sequencing in environmental microorganisms research

摘要:

1975年，第一个cDNA的基因组——噬菌体фX174基因组的测序完成，标志着测序时代的开始。1996年以来人类基因组计划的发起极大地推动了测序技术的应用和发展，第二代测序技术也被称为高通量测序技术。而单分子DNA测序技术是最近10年内发展起来的新一代的测序技术，称为第三代测序技术，其中包括单分子实时测序(Single molecule real time sequencing，SMRT)、真正单分子测序和单分子纳米孔测序等技术。SMRT测序技术有超长读长、测序周期短、不需要模板扩增和直接检测表观修饰位点等特点，为研究人员提供了新的选择。本文综述了SMRT测序技术的基本原理、性能以及它在微生物16S rRNA基因、微生物全基因组以及微生物宏基因组测序等方面的应用，并分析了SMRT测序技术在环境微生物应用中的优势以及存在的问题。

Abstract:

In 1975, the sequencing of the first cDNA genome, the phage фX174 genome, marked the beginning of the sequencing era. Since 1996, the launch of the Human Genome Project has greatly promoted the application and development of sequencing technology. The second generation sequencing technology is also known as high-throughput sequencing technology. Single-molecule DNA sequencing technology is a new generation of sequencing technology called third-generation sequencing technology, which developed in the last 10 years. It includes single-molecule real-time sequencing, true single-molecule sequencing and single-molecule nanopore sequencing. Because SMRT sequencing technology has the characteristics of long reading length, short sequencing period, without requirement for template amplification and direct detection of apparent modification sites, it provides researchers with new options. This paper reviews the basic principle and performance of SMRT sequencing technology and its application in microbial 16S rRNA gene, microbial whole genome sequencing and microbial macrogenome sequencing, and analyzes the advantages and problems of SMRT sequencing technology in environmental microorganisms.