Misannotations of rRNA can now generate 90% false positive protein matches in metatranscriptomic studies.

Misannotations of rRNA can now generate 90% false positive protein matches in metatranscriptomic studies.
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DOI:
10.1093/nar/gkr576
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发表时间:
2011-11-01
影响因子:
14.9
通讯作者:
Zehr JP
Zehr JP
中科院分区:
生物学2区
文献类型:
--
作者:
Tripp HJ;Hewson I;Boyarsky S;Stuart JM;Zehr JP

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在分析来自西南太平洋和赤道大西洋的23个站点的9522746个焦磷酸测序读数的过程中,我们注意到rRNA作为蛋白质的错误注释现在是如此普遍,以至于rRNA焦磷酸测序读数与美国国家生物技术信息中心(NCBI)非冗余蛋白质数据库的假阳性匹配接近90%。23 S rRNA的一个保守部分经常被错误注释,足以促使Pfam的管理员创建一个虚假的蛋白质家族。对伪Pfam蛋白家族(PF 10695,“Cw-水解酶”)中每个种子序列的注释历史的详细检查发现了主要测序中心的标准操作程序和质量保证程序中的问题,以及与管理公共数据库(如GenBank和SwissProt)的管理实践有关的其他问题。我们为所有这些问题提供了建议,并建议元转录组学领域的工作人员格外小心,避免在他们的数据集中包含假阳性匹配。
In the course of analyzing 9 522 746 pyrosequencing reads from 23 stations in the Southwestern Pacific and equatorial Atlantic oceans, it came to our attention that misannotations of rRNA as proteins is now so widespread that false positive matching of rRNA pyrosequencing reads to the National Center for Biotechnology Information (NCBI) non-redundant protein database approaches 90%. One conserved portion of 23S rRNA was consistently misannotated often enough to prompt curators at Pfam to create a spurious protein family. Detailed examination of the annotation history of each seed sequence in the spurious Pfam protein family (PF10695, ‘Cw-hydrolase’) uncovered issues in the standard operating procedures and quality assurance programs of major sequencing centers, and other issues relating to the curation practices of those managing public databases such as GenBank and SwissProt. We offer recommendations for all these issues, and recommend as well that workers in the field of metatranscriptomics take extra care to avoid including false positive matches in their datasets.
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