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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Sequencing and characterization of Salmonella typhi plasmid R27 (incompatibility group HI1) trhC, a transfer gene encoding a potential nucleoside triphosphate-binding domain.
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Sequencing and characterization of Salmonella typhi plasmid R27 (incompatibility group HI1) trhC, a transfer gene encoding a potential nucleoside triphosphate-binding domain.

机译：鼠伤寒沙门氏菌质粒R27（不相容组HI1）trhC的测序和鉴定，trhC是编码潜在的核苷三磷酸结合域的转移基因。

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摘要

IncHI1 plasmids are one of the few plasmids known to mediate multiple antibiotic resistance in Salmonella typhi. These plasmids are temperature-sensitive for transfer and R27 is the prototype plasmid. DNA sequencing within the Tra2 region of R27, encoding genes involved in mating pair formation, identified trhC encoding the TrhC protein of 90,000 Da, which was visualized using an in vitro transcription/translation system. Expression of the TrhC protein also identified two smaller protein products of approximately 23 and 25 kDa which are predicted to be protease digestion products. The migration of these smaller products changed when the reactions were run at 28 vs 37 degrees C. The TrhC protein contained a putative nucleotide triphosphate-binding protein and exhibited sequence similarities with several other proteins implicated in bacterial conjugation, including TraC (F), TraB (pKM101), VirB4 (Ti), TrbE (RP4). Phylogenetic analysis showed TrhC was most closely related to TraC. Mini-Tn10 insertions in trhC generated transfer defective mutants, and no pili were specified by the trhC mutants. The trhC gene appeared to be a hot spot for transposon insertion as 37.5% mapped into this open reading frame. One trhC mutant, pDT2990, was able to be complemented by a cloned trhC gene giving a transfer frequency of 1 x 10(-3) transconjugants per recipient in an 18-h mating, whereas the wild-type transfer frequency of R27 was 1 x 10(-2) transconjugants per recipient. Copyright 1999 Academic Press.
机译：IncHI1质粒是已知在伤寒沙门氏菌中介导多种抗生素抗性的少数质粒之一。这些质粒对转移温度敏感，R27是原型质粒。 R27的Tra2区域内的DNA测序（编码参与配对形成的基因）鉴定出编码90,000 Da的TrhC蛋白的trhC，可使用体外转录/翻译系统将其可视化。 TrhC蛋白的表达还鉴定出了大约23和25 kDa的两个较小的蛋白质产物，预计它们是蛋白酶消化产物。当反应在28 vs 37摄氏度下进行时，这些较小产物的迁移发生了变化。TrhC蛋白包含一个推定的核苷酸三磷酸结合蛋白，并且与涉及细菌结合的其他几种蛋白（包括TraC（F），TraB）具有序列相似性（pKM101），VirB4（Ti），TrbE（RP4）。系统发育分析表明TrhC与TraC关系最密切。 trhC中的Mini-Tn10插入会产生转移缺陷突变体，并且trhC突变体未指定菌毛。 trhC基因似乎是转座子插入的热点，因为有37.5％的基因定位在此开放阅读框中。一个trhC突变体pDT2990能够与克隆的trhC基因互补，在18小时的交配中每个受体的转移频率为1 x 10（-3）个转导结合体，而R27的野生型转移频率为1 x每个收件人10（-2）个转导结合剂。版权所有1999 Academic Press。

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