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通过直接PCR分析检测发现,空肠弯曲菌染色体DNA中的同核苷酸序列呈现出高频多态性。

Homonucleotide stretches in chromosomal DNA of Campylobacter jejuni display high frequency polymorphism as detected by direct PCR analysis.

作者信息

Wassenaar Trudy M, Wagenaar Jaap A, Rigter Alan, Fearnley Cathrine, Newell Diane G, Duim Birgitta

机构信息

Molecular Microbiology and Genomics Consultants, Tannenstrasse 7, D-55576 Zotzenheim, Germany.

出版信息

FEMS Microbiol Lett. 2002 Jun 18;212(1):77-85. doi: 10.1111/j.1574-6968.2002.tb11248.x.

Abstract

Homopolymeric nucleotide tracts have been previously identified in the genome sequence of Campylobacter jejuni 11168 [Parkhill et al., Nature 403 (2000) 665-668]. These tracts are believed to regulate contingency genes but as yet no phenotypic variation has been identified associated with many of these genes. To investigate homopolymeric tracts for genes for which there is no observable phenotype, a method was designed to visualise profiles of the various tract lengths directly at the genomic level by means of PCR and denatured polyacrylamide gel electrophoresis. Six of the seven contingency genes investigated displayed variation in the length of the respective homonucleotide tracts. Surprisingly, each contingency gene gave a typical peak profile that represented a conserved size distribution of polymorphic forms. For each gene studied, peak profiles were conserved between strains of C. jejuni. Duplicated genes, containing homonucleotide stretches, displayed locus-specific peak distributions for each gene copy. Contingency genes were polymorphic within single colonies, and the observed complex peak profiles suggested a frequency of slippage several orders of magnitude higher than reported for other organisms. No G7 (or C7) stretch was ever observed, and their absence from the complete genome suggests strong selection against their presence. In view of the predictable outcome of the process leading to these polymorphisms, it is hypothesised that the formation and/or selection of these tracts is not a random process, but is driven by as yet unknown mechanism(s). High-frequency polymorphism of these genes may be a mechanism by which C. jejuni survives selection bottlenecks between opportunities for growth within a host.

摘要

先前已在空肠弯曲菌11168的基因组序列中鉴定出同聚核苷酸序列[帕克希尔等人,《自然》403(2000)665 - 668]。这些序列被认为可调控应急基因,但迄今为止,尚未发现与许多此类基因相关的表型变异。为了研究那些没有可观察到表型的基因的同聚序列,设计了一种方法,通过聚合酶链反应(PCR)和变性聚丙烯酰胺凝胶电泳,直接在基因组水平上可视化各种序列长度的图谱。所研究的7个应急基因中有6个在各自同核苷酸序列的长度上表现出变异。令人惊讶的是,每个应急基因都给出了一个典型的峰图谱,代表了多态形式的保守大小分布。对于所研究的每个基因,空肠弯曲菌菌株之间的峰图谱是保守的。含有同核苷酸延伸的重复基因,每个基因拷贝显示出基因座特异性的峰分布。应急基因在单个菌落内具有多态性,观察到的复杂峰图谱表明其滑动频率比其他生物体报道的高几个数量级。从未观察到G7(或C7)序列延伸,并且它们在完整基因组中的缺失表明对其存在有强烈的选择压力。鉴于导致这些多态性的过程的可预测结果,推测这些序列的形成和/或选择不是一个随机过程,而是由尚未知的机制驱动的。这些基因的高频多态性可能是空肠弯曲菌在宿主内生长机会之间的选择瓶颈中生存的一种机制。

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