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Analyses of amber mutants of Salmonella phage SPN3US.

机译：沙门氏菌噬菌体SPN3US的琥珀色突变体的分析。

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摘要

The giant Salmonella enterica phage SPN3US belongs to the &phis;KZ-related phages which are amongst the most complex virions of prokaryotic viruses, even more complex than most eukaryotic viruses. SPN3US for example assigns 46% of its 240kbp genome to code for virion proteins. We have limited knowledge about these giant phages as they encode extremely diverged genes, from other phages and even to one another. The &phis;KZ-related phages possess large genomes and correspondingly large and complex capsids, two sets of fragmented RNAP beta and beta'-like proteins and they also exhibit an unusual replication having a DNA polymerase split into two polypeptides.;The complete genome of phage SPN3US was previously sequenced by Lee et al. (2011). As is typical for a newly sequenced phage, 80% of its ORFs were annotated as hypothetical. To address this problem of functionally unassigned genes, a novel strategy for phages is to target the essential genes and work to determine their functions using genetic approaches. In this work amber mutants of a giant phage were sequenced for the first time. Once the amber mutation sites were determined the sequence similarity of those genes in other &phis;KZ-related phages was analyzed. This has enabled us to identify fourteen essential genes in SPN3US, ten of which code for the components of the phage virion. Previously we knew the function of three genes, now we can infer the function of four more genes. The mutants encoding these essential genes will now form the basis of further studies to determine specific functions for these novel genes. Ten of these genes are well conserved among the &phis;KZ-related phages being classified as core genes for this subfamily.;These studies have demonstrated that SPN3US is an excellent model system to study the &phis;KZ-related phages.
机译：巨大的肠炎沙门氏菌噬菌体SPN3US属于与KZ相关的噬菌体，它们是原核病毒最复杂的病毒体之一，甚至比大多数真核病毒更复杂。例如，SPN3US将其240kbp基因组的46％分配给病毒粒子蛋白。我们对这些巨大的噬菌体了解甚少，因为它们编码极为不同的基因，从其他噬菌体到彼此。与KZ有关的噬菌体具有大的基因组和相应的大而复杂的衣壳，两组片段化的RNAP beta和beta'样蛋白，并且它们还表现出不寻常的复制，其DNA聚合酶被分为两个多肽。噬菌体SPN3US先前由Lee等人测序。 （2011）。正如新测序的噬菌体的典型情况一样，其80％的ORF都被注释为假设的。为了解决功能上未分配的基因的问题，噬菌体的新策略是靶向必需基因并使用遗传方法来确定其功能。在这项工作中，首次对巨噬细胞的琥珀突变体进行了测序。一旦确定了琥珀色突变位点，就分析了这些与其他与KZ相关的噬菌体中的基因的序列相似性。这使我们能够鉴定SPN3US中的14个必需基因，其中10个编码噬菌体病毒体的成分。以前我们知道三个基因的功能，现在我们可以推断出另外四个基因的功能。现在，编码这些必需基因的突变体将构成进一步研究的基础，以确定这些新基因的特定功能。这些基因中有10个在与KZ相关的噬菌体中被保守为该亚科的核心基因。这些研究表明SPN3US是研究与KZ相关的噬菌体的优秀模型系统。

著录项

  • 作者

    Benitez Quintana, Andrea Denisse.;

  • 作者单位

    Rochester Institute of Technology.;

  • 授予单位 Rochester Institute of Technology.;
  • 学科 Bioinformatics.;Microbiology.;Molecular biology.
  • 学位 M.S.
  • 年度 2016
  • 页码 96 p.
  • 总页数 96
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类 公共建筑;
  • 关键词

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