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首页> 外文期刊>Infection and immunity >Use of transposon Tn916 to inactivate and isolate a mutacin-associated gene from Streptococcus mutans.
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Use of transposon Tn916 to inactivate and isolate a mutacin-associated gene from Streptococcus mutans.

机译：使用转座子Tn916灭活并从变形链球菌中分离出与诱变素相关的基因。

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Among the attributes thought to contribute to the virulence of Streptococcus mutans is its ability to elaborate bacteriocinlike substances, which may provide a selective force enhancing its colonization potential. One such inhibitory substance, mutacin II, is produced by certain plasmid-containing strains of S. mutans. We introduced insertional mutations into a mutacin II-producing strain of S. mutans (UA96) by transformation with a plasmid carrying Tn916, resulting in transformants bearing single inserts of the transposon at different sites within the chromosome. The insertions identify five different EcoRI fragments required for production of mutacin II (Bac phenotype; bac-1 to bac-5 genotypes). The EcoRI fragments, containing bac-1::Tn916 was ligated into a cosmid vector, pJC74, and transduced into Escherichia coli DH1, where Tn916 is known to be unstable. The loss of Tn916 resulted in a 30-kb plasmid, pPC974, containing approximately 15 kb of S. mutans DNA. A Bac-associated DNA fragment was then subcloned into the streptococcus-E. coli shuttle vector pVA838 and transformed into S. mutants, where it was capable of complementing the bac mutation in the Bac- parent. These findings suggest that we have isolated at least one gene associated with mutacin production.
机译：被认为对变形链球菌的毒性有贡献的特性之一是它具有精心设计的细菌素样物质的能力，这可以提供增强其定殖潜力的选择性力量。一种这样的抑制物质，诱变蛋白II，是由某些含有质粒的变形链球菌菌株产生的。我们通过用携带Tn916的质粒进行转化，将插入突变引入产生诱变酶II的变形链球菌（UA96）菌株中，导致转化子在染色体内的不同位点带有转座子的单个插入片段。该插入物鉴定了产生诱变蛋白II（Bac表型; bac-1至bac-5基因型）所需的五个不同的EcoRI片段。将含有bac-1 :: Tn916的EcoRI片段连接到粘粒载体pJC74中，并转导到大肠杆菌DH1中，其中Tn916不稳定。 Tn916的缺失导致了一个30 k​​b的质粒pPC974，其中包含约15 kb的变形链球菌DNA。然后将与Bac相关的DNA片段亚克隆到链球菌E中。大肠杆菌穿梭载体pVA838并转化为S.突变体，在其中它能够补充Bacparent中的bac突变。这些发现表明我们已经分离出至少一种与诱变素产生有关的基因。

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