基于SMRT测序技术的16S rRNA基因全长测序及其分析方法  

Full-length Sequencing of 16S rRNA Gene and Its Analysis Based on the SMRT Sequencing Technology

作　　者：唐勇[1,2] 刘旭 

机构地区：[1]乐山职业技术学院,乐山614000 [2]乐山丰野农业科技有限责任公司,乐山614000 [3]乐山市农业局,乐山614000

出　　处：《生物技术通报》2017年第8期34-39,共6页Biotechnology Bulletin

基　　金：乐山市科技局重点研究项目(16JSC128)

摘　　要：被称为第三代测序技术的单分子测序是最近几年发展起来的高通量测序技术。其中,由Pacbio Bio Sciences公司开发的单分子实时测序技术(SMRT)是最先商用的技术。SMRT测序技术通过对模板序列循环测序产生环形一致序列(CCS),成功克服第三代测序技术准确率低的弊病。通过SMRT测序技术,科学家可以更深入准确地探究复杂环境微生物的结构和功能。介绍SMRT测序技术在微生物16S rRNA基因测序中的优势和劣势,并就基于SMRT测序技术所得的全长16S rRNA基因序列的质量控制、错误序列排除、聚类和注释分析等重要分析环节进行概述,同时,提出利用SMRT测序技术研究复杂环境微生物可能存在的问题及其解决方法,期望能为研究人员提供参考。Single-molecule sequencing,called as third-generation sequencing technology,is a high-throughput technique developedin last few years. Of them,single-molecule real-time（SMRT）sequencing technology,developed by Pacific Bio Sciences（Pac Bio）,is thefirst commercial technology. SMRT sequencing technology could successfully overcome the disadvantage of low accuracy in the third-generationsequencing technology,by generating circular consensus sequence（CCS）through cycle sequencing the template sequence. Therefore,SMRT sequencing technology will allow scientists to profoundly and accurately study the structures and functions of microbial communities incomplex environment. Here,we introduced the advantages and disadvantages of SMRT sequencing technology in 16 S rRNA gene sequence ofmicroorganism,and summarized the important steps,such as quality control,filtering of error tags,clustering analysis,annotation analysis,etc. of full-length 16 S rRNA gene sequence acquired by SMRT sequencing technology. In addition,we pointed out the problems and thefeasible solutions while applying SMRT sequencing technology in the study of microbial in complex environment,aiming at providing referencesfor researchers in this field.

关 键 词：单分子实时测序技术 PAC BIO RSⅡ 第三代测序技术 环形一致序列 

分 类 号：Q78[生物学—分子生物学]