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应用 Solexa 深度测序和生物信息学分析技术研究具有重要公共卫生意义的带科绦虫微小 RNA。

Characterization of microRNAs in Taenia saginata of zoonotic significance by Solexa deep sequencing and bioinformatics analysis.

机构信息

National Institute of Parasitic Diseases, Chinese Center for Disease Control and Prevention, Laboratory of Parasite and Vector Biology, Ministry of Health, WHO Collaborating Center for Malaria, Schistosomiasis and Filariasis, Shanghai, 200025, Peoples Republic of China.

出版信息

Parasitol Res. 2012 Jun;110(6):2373-8. doi: 10.1007/s00436-011-2773-x. Epub 2011 Dec 28.

Abstract

The beef tapeworm Taenia saginata infects human beings with symptoms ranging from nausea, abdominal discomfort to digestive disturbances and intestinal blockage. In the present study, microRNA (miRNA) expressing profile in adult T. saginata was analyzed using Solexa deep sequencing and bioinformatics analysis. A total of 15.8 million reads was obtained by Solexa sequencing, and 13.3 million clean reads (1.73 million unique sequences) was obtained after removing reads smaller than 18 nt. Ten conserved miRNAs corresponding to 607,382 reads were found when matching the reads against known miRNAs of Schistosoma japonicum in miRBase database. The miR-71 had the most abundant expression in T. saginata, followed by miR-219-5p, but some other common miRNAs such as let-7, miR-40, and miR-103 were not identified in T. saginata. Nucleotide bias analysis found that the known miRNAs showed high bias and the uracil was the dominant nucleotide, particularly at the first and 11th positions which were almost at the beginning and middle of conserved miRNAs. One novel miRNA (Tsa-miR-001) corresponding to ten precursors was identified and confirmed by stem-loop RT-PCR. To our knowledge, this is the first report of miRNA profiles in T. saginata, which will contribute to better understanding of the complex biology of this zoonotic trematode. The reported data of T. saginata miRNAs should provide valuable references for miRNA studies of closed related zoonotic Taenia cestodes such as Taenia solium and Taenia asiatica.

摘要

牛带绦虫 Taenia saginata 感染人体后可引起恶心、腹部不适、消化功能紊乱和肠梗阻等症状。本研究采用高通量测序和生物信息学分析方法,对牛带绦虫成虫的 microRNA(miRNA)表达谱进行了分析。Solexa 测序共获得 1580 万条读段,去除小于 18nt 的读段后,共获得 1330 万条清洁读段(173 万条独特序列)。将这些读段与 miRBase 数据库中日本血吸虫的已知 miRNA 进行比对,共发现 10 个保守 miRNA,对应 607382 条读段。牛带绦虫中 miR-71 的表达丰度最高,其次是 miR-219-5p,但 miR-40、miR-103 等一些常见 miRNA 未在牛带绦虫中检测到。核苷酸偏好性分析发现,已知 miRNA 表现出高度的偏好性,且尿嘧啶是主要的核苷酸,尤其是在保守 miRNA 的第 1 位和第 11 位,这两个位置几乎位于 miRNA 的起始和中间位置。通过茎环 RT-PCR 鉴定并验证了一个对应于 10 个前体的新 miRNA(Tsa-miR-001)。据我们所知,这是首次对牛带绦虫 miRNA 进行的研究,有助于更好地了解这种人畜共患的吸虫的复杂生物学。本研究报道的牛带绦虫 miRNA 数据可为密切相关的人畜共患带绦虫(如猪带绦虫和亚洲带绦虫)的 miRNA 研究提供有价值的参考。

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