摘要
高通量测序技术是近年来发展起来的一种重要的分子生物学技术,能一次对几十万至几百万条DNA分子进行序列测定,在基因组解析、转录组分析、基因变异位点筛查等方面广泛应用。探讨通过对感染病毒的草莓材料进行转录组测序来解析病毒基因组的可行性。以怀疑感染病毒的草莓叶片为试材,利用Illumina测序技术进行转录组测序分析。通过对组装得到的单基因序列进行注释分析,筛选出12个注释为草莓病毒的单基因序列,它们源自4种草莓病毒,分别是草莓轻型黄边病毒、草莓坏死休克病毒、草莓镶脉病毒及草莓皱缩病毒。利用RT-PCR技术对转录组测序结果进行了验证,表明利用高通量测序技术解析草莓病毒基因组序列的可靠性。本研究探索出解析草莓病毒基因组序列信息的新方法,为研究草莓病毒的进化和病毒分子检测奠定基础。
Recently, high-throughput sequencing, one of the most important molecular biology technology that can sequence hundreds of thousands to millions DNA molecule once, has a wide range of applications in genome analysis, transcriptome analysis, screening of gene mutation sites. In this study, the feasibility of determination of virus genomic sequence by transcriptome assembly from RNA-Seq data of strawberry plants infected with virus was explored. The strawberry leaves suspected to be infected with viruses were used as materials for transcriptome sequencing with Illumina method. Twelve unigenes annotated strawberry viruses were filtered out by annotating the assembled unigenes, and they belong to Strawberry mild yellow edge virus,Strawberry necrotic shock virus, Strawberry vein banding virus and Strawberry crinkle virus. The result of RT-PCR indicated that it was reliability to determine the genomic sequences of strawberry viruses through RNA-Seq. This study explores a new method for determining the genomic sequences of strawberry viruses, which lays the foundation for the research on the evolution and detection of strawberry viruses.
出处
《沈阳农业大学学报》
CSCD
北大核心
2016年第5期536-540,共5页
Journal of Shenyang Agricultural University
基金
国家自然科学基金项目(31470678)
