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首页> 外文期刊>Folia Microbiologica >Identification of a holin encoded by theStreptomyces aureofaciens phage µ1/6; functional analysis inEscherichia coli system
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Identification of a holin encoded by theStreptomyces aureofaciens phage µ1/6; functional analysis inEscherichia coli system

机译：鉴定出由金黄色链霉菌噬菌体μ1/ 6编码的霍林；大肠杆菌系统功能分析

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摘要

An open reading frame encoding an 88 amino acid protein was present downstream of the previously characterized endolysin ofStreptomyces aureofaciens phage µ1/6. Structural analysis of its sequence revealed features characteristic for holin. This open reading frame encoding the putative holin was amplified by polymerase chain reaction and cloned into the expression vector pET-21d(+). Synthesis of the holin-like protein resulted in bacterial cell death but not lysis. Theholµ1/6 gene was able to complement the defective λS allele in the nonsuppressingEscherichia coli HB101 strain to produce phage progeny. This fact suggests that the proteins encoded by both phage genes have analogous function,i.e. the streptomycete holin induces nonspecific lesions in the cytoplasmic membrane, through which the λ endolysin gains an access to its substrate, the cell wall. The concomitant expression of bothS. aureofaciens holµ1/6 and λ endolysin inE. coli resulted in abrupt cell lysis. This result provided further evidence that the product ofholµ1/6 gene is a holin.
机译：在先前表征的金黄色链霉菌噬菌体μ1/ 6的内溶素下游存在编码88个氨基酸蛋白的开放阅读框。其序列的结构分析揭示了霍林蛋白的特征。通过聚合酶链反应扩增编码推定的holin的开放阅读框，并将其克隆到表达载体pET-21d（+）中。霍林样蛋白的合成导致细菌细胞死亡，但不裂解。 holµ1 / 6基因能够补充非抑制性大肠杆菌HB101菌株中有缺陷的λS等位基因，从而产生噬菌体后代。这个事实表明由两个噬菌体基因编码的蛋白质具有类似的功能，即。链霉菌素holin诱导细胞质膜中的非特异性损伤，λ溶素通过该损伤进入其底物即细胞壁。 bothS的伴随表达。金黄色葡萄球菌holµ1 / 6和λ内溶素inE。大肠杆菌导致细胞突然裂解。该结果提供了进一步的证据，证明holμ1/ 6基因的产物是holin。

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  • 来源
    《Folia Microbiologica》 |2004年第6期|679-684|共6页
  • 作者

    J. Farkašovská; A. Godány; C. Vlček;

  • 作者单位

    Institute of Molecular Biology Slovak Academy of Sciences;

    Institute of Molecular Biology Slovak Academy of Sciences;

    Center for Integrated Genomics Institute of Molecular Genetics Academy of Sciences of the Czech Republic;

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