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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Characterization of a cryptic plasmid pSFKW33 from Shewanella sp. 33B.
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Characterization of a cryptic plasmid pSFKW33 from Shewanella sp. 33B.

机译：来自希瓦氏菌属的隐性质粒pSFKW33的表征。 33B。

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摘要

A cryptic plasmid pSFKW33 from psychrotrophic bacterium Shewanella sp. 33B, an isolate from the Gulf of Gdansk (the Baltic Sea), was sequenced and characterized. It is an 8021bp circular molecule with 38% GC content, which shows a distinctive nucleotide sequence without homology to other known plasmids. The nucleotide sequence analysis predicts eight open reading frames. The deduced amino acid sequence of ORF-1 shared significant similarity with the phage integrase protein from Pseudomonas putida GB-1. The ORF2 product showed some similarity with the hypothetical protein Shal_1786 from Shewanella halifaxensis HAW-EB4. The ORF-3 product revealed high amino acid sequence homology with the Escherichia coli replication initiation protein similar to pi protein of R6K plasmid. Five repeat regions (three perfect 22-bp repeats and two imperfect motifs), a putative ribosome binding site, and -10 and -35 promotor sequences were identified upstream of the ORF-3 (rep). The rep module is very similar to several theta replicating iteron family plasmids, suggesting that pSFKW33 replication follows the same course. The ORF-5 product revealed significant identity with the entry exclusion protein 1 of plasmid pIS2 from enteropathogenic strain E. coli 0111:H. The ORF-6 product showed significant identity with the putative transcriptional regulator protein from Shewanella benthica KT99 belonging to HTH-XRE-family protein. The ORF-8 encoded protein that showed some similarity with the hypothetical protein KT99_00146 from Shewanella benthica KT99. The ORF-4 and ORF-7 encoded putative proteins of 189 and 118 amino acid residues, respectively, with no homologies to any known proteins. The absolute copy number of pSFKW33 was estimated to be five copies per chromosome by real-time PCR.
机译：来自精神营养细菌Shewanella sp。的隐性质粒pSFKW33。对来自格但斯克湾（波罗的海）的分离株33B进行了测序和鉴定。它是一个8021bp的环状分子，GC含量为38％，显示出独特的核苷酸序列，与其他已知质粒无同源性。核苷酸序列分析预测了八个开放阅读框。推导的ORF-1氨基酸序列与恶臭假单胞菌GB-1的噬菌体整合酶蛋白具有显着相似性。 ORF2产物与来自希瓦氏菌HAW-EB4的假设蛋白Shal_1786有相似之处。 ORF-3产物显示出与R6K质粒的pi蛋白相似的大肠杆菌复制起始蛋白的高氨基酸序列同源性。在ORF-3（rep）的上游发现了五个重复区域（三个完美的22 bp重复和两个不完美的基序），一个假定的核糖体结合位点以及-10和-35启动子序列。 rep模块与几个theta复制iteron家族质粒非常相似，表明pSFKW33复制遵循相同的过程。 ORF-5产物与肠道致病菌株大肠杆菌0111：H的质粒pIS2的进入排斥蛋白1具有显着同一性。 ORF-6产物与Shehtella benthica KT99的推测的转录调节蛋白具有明显的同一性，属于HTH-XRE家族蛋白。 ORF-8编码的蛋白质与来自Shewanella benthica KT99的假设蛋白质KT99_00146表现出一些相似性。 ORF-4和ORF-7分别编码189和118个氨基酸残基的推定蛋白质，与任何已知蛋白质均无同源性。通过实时PCR，pSFKW33的绝对拷贝数估计为每个染色体5个拷贝。

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