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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Genetic and transcriptional analysis of a regulatory region in streptococcal conjugative transposon Tn5252.
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Genetic and transcriptional analysis of a regulatory region in streptococcal conjugative transposon Tn5252.

机译：链球菌共轭转座子Tn5252调控区的遗传和转录分析。

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摘要

In an attempt to increase our understanding of the mechanisms of conjugal transposition among gram-positive bacteria, we analyzed the genetic and structural properties of a 1.2-kb DNA fragment at the left end of the streptococcal conjugative transposon Tn5252. The sequence data revealed four short open reading frames. Polypeptides likely to correspond to two of these genes were identified. Transcriptional start sites and the promoter sequences of three transfer-related genes in the left terminal region of the element were identified. The deduced amino acid sequence of one of these, ORF3, was found to be similar to that of several prokaryotic transcriptional regulator proteins. Insertion mutagenesis at this locus reduced the transfer of the element by three orders of magnitude. The presence of a multicopy plasmid carrying ORF3 in a donor cell carrying Tn5252 with a mutated copy of ORF3 or an unaltered element also reduced the transfer frequency of the element similarly. Gel mobility shift assays showed that the ORF3 protein was capable of binding to not only other discrete sites at the left end of the element but also its own promoter, suggesting autoregulation. These results indicate that the ORF3 protein is involved in the regulation of the conjugative transposition of the element. Copyright 2000 Academic Press.
机译：为了增加我们对革兰氏阳性细菌之间接合转座机制的了解，我们分析了链球菌接合转座子Tn5252左端1.2kb DNA片段的遗传和结构特性。序列数据揭示了四个短的开放阅读框。确定了可能对应于这些基因中的两个的多肽。确定了元件左端区域中三个转移相关基因的转录起始位点和启动子序列。发现其中之一ORF3的推导氨基酸序列与几种原核转录调节蛋白的推导氨基酸序列相似。在该基因座处的插入诱变将元素的转移减少了三个数量级。类似地，在携带带有突变拷贝的ORF3或未改变元件的Tn5252的供体细胞中，存在携带ORF3的多拷贝质粒也降低了该元件的转移频率。凝胶迁移率迁移分析表明，ORF3蛋白不仅能够与元件左端的其他离散位点结合，还能够与自身的启动子结合，从而提示自身调节。这些结果表明ORF3蛋白参与元件的共轭转座的调节。版权所有2000学术出版社。

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