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首页> 外文期刊>Plasmid: An International Journal Devoted to Extrachromosomal Gene Systems >Molecular characterization of a cryptic plasmid from Campylobacter jejuni.
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Molecular characterization of a cryptic plasmid from Campylobacter jejuni.

机译：空肠弯曲杆菌隐性质粒的分子表征。

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摘要

Campylobacter jejuni is a leading bacterial cause of human enterocolitis. Molecular genetic characterization of this pathogen has been hampered by the lack of genetic tools that are functional in this organism. Cloning vectors commonly used in other organisms usually do not replicate within C. jejuni. To develop a system for functional analysis of C. jejuni genes, a small plasmid (pCJ01) identified in a poultry isolate of C. jejuni was sequenced and characterized in this study. By using inverse PCR, the full sequence of pCJ01 was amplified and subsequently determined. Results indicate that pCJ01 is a circular molecule of 3212 bp, with a G + C content of 33.5%. A typical plasmid replication origin with iteron sequences is identified upstream of the DNA sequences encoding replication initiation proteins. Four open reading frames (ORFs) are present in pCJ01. ORF1 and ORF2 share high homology with the putative RepA and RepB proteins, respectively, of known C. coli plasmids. ORF3 and ORF4, of unknown function, do not exhibit homology with any sequences deposited in the GenBank database. Hydropathy analysis predicts that ORF3 and ORF4 contain multiple stretches of hydrophobic amino acids, suggesting that they may encode transmembrane proteins. Since pCJ01 is a small plasmid and can be readily prepared from C. jejuni, it may be modified for use in molecular characterization of C. jejuni virulence genes. Copyright 2001 Academic Press.
机译：空肠弯曲菌是人小肠结肠炎的主要细菌病因。该病原体的分子遗传学特征已因缺乏在该生物体中起作用的遗传工具而受到阻碍。通常在其他生物中使用的克隆载体通常不会在空肠弯曲杆菌内复制。为了开发用于空肠弯曲杆菌基因功能分析的系统，本研究对空肠弯曲杆菌家禽分离物中鉴定出的小质粒（pCJ01）进行了测序和鉴定。通过反向PCR，扩增并随后确定pCJ01的完整序列。结果表明pCJ01是3212 bp的环状分子，G + C含量为33.5％。在编码复制起始蛋白的DNA序列的上游鉴定出具有iteron序列的典型质粒复制起点。 pCJ01中有四个开放阅读框（ORF）。 ORF1和ORF2与已知C.coli质粒的推定RepA和RepB蛋白分别具有高度同源性。功能未知的ORF3和ORF4与GenBank数据库中存放的任何序列均不具有同源性。亲水性分析预测ORF3和ORF4包含多个疏水性氨基酸片段，表明它们可能编码跨膜蛋白。由于pCJ01是一种小质粒，可以很容易地从空肠弯曲杆菌制备，因此可以对其进行修饰，以用于空肠弯曲杆菌毒力基因的分子鉴定。版权所有2001学术出版社。

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