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首页> 外文期刊>Infection and immunity >Analysis of the Organization of Multicopy Linear- and Circular-Plasmid-Carried Open Reading Frames inBorrelia burgdorferi Sensu Lato Isolates
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Analysis of the Organization of Multicopy Linear- and Circular-Plasmid-Carried Open Reading Frames inBorrelia burgdorferi Sensu Lato Isolates

机译：伯氏疏螺旋体中多拷贝线性和圆形质粒携带的开放阅读框的组织分析

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摘要

Plasmid cp8.3 of Borrelia afzelii IP21 carries several open reading frames (ORFs) and a 184-bp inverted repeat (IR) element. It has been speculated that this plasmid may encode factors involved in virulence or infectivity. In this report, we have characterized the distribution, molecular variability, and organization of ORFs 1, 2, and 4 and the IR elements among isolates of the Borrelia burgdorferi sensu lato complex. ORFs 1 and 2 are contained within a segment of cp8.3 that is bordered by the IR elements, while ORF 4 resides just outside of the IR-bordered region. By PCR, ORF 4 was amplified from most isolates while ORFs 1 and 2 were amplified from only some B. afzelii isolates. However, Southern hybridization analyses with ORF 1, 2, and 4 probes detected related sequences even in some isolates that were PCR negative. The ORF restriction fragment length polymorphism patterns varied widely even among isolates of the same species. Two-dimensional contour-clamped homogeneous electric field–pulsed-field gel electrophoresis and Southern hybridization detected ORF 1-, 2-, and 4-related sequences on linear and circular plasmids. In addition, an ORF 4-related sequence was detected on a previously uncharacterized, circular plasmid that is greater than 70 kb in size. The IR elements originally identified on plasmid cp8.3 of B. afzelii IP21 were also analyzed by Southern hybridization. Related sequences were detected in some but not all B. burgdorferi sensu lato isolates. These sequences are carried on plasmids in addition to cp8.3 in some isolates. Single-primer PCR analyses demonstrated that in some isolates these sequences exist with IR orientation. The data presented here demonstrate that the IR elements and the ORF 1-, 2-, and 4-related sequences are multicopy and are variable in organization and in genomic location among isolates of the B. burgdorferi sensu lato complex. These analyses provide additional evidence for the highly variable organization of the plasmid component of the B. burgdorferi sensu lato genome.
机译：短孢单胞菌IP21的质粒cp8.3带有几个开放阅读框（ORF）和一个184 bp反向重复序列（IR）元件。已经推测该质粒可以编码涉及毒力或感染性的因子。在本报告中，我们表征了 Borrelia burgdorferi sensu lato复杂菌株中ORF 1、2和4的分布，分子变异性和组织以及IR元素。 ORF 1和2包含在以IR元素为边界的cp8.3片段中，而ORF 4恰好位于IR边界区域之外。通过PCR，从大多数分离物中扩增出ORF 4，而仅从某些 B中扩增出ORF 1和2。 afzelii 分离株。但是，使用ORF 1、2和4探针进行的Southern杂交分析甚至在某些PCR阴性的分离物中也检测到相关序列。即使在相同物种的分离物中，ORF限制性片段长度多态性模式也存在很大差异。二维轮廓固定的均相电场-脉冲场凝胶电泳和Southern杂交检测线性和环状质粒上的ORF 1、2和4相关序列。另外，在先前未表征的环状质粒中检测到ORF 4相关序列，其大小大于70 kb。最初在 B的质粒cp8.3上鉴定的IR元件。还通过Southern杂交分析了afzelii IP21。在一些但不是全部 B中检测到相关序列。 burgdorferi sensu lato分离株。这些序列在某些分离株中除了cp8.3外还携带在质粒上。单引物PCR分析表明，在某些分离物中，这些序列以IR方向存在。此处提供的数据表明，IR元件以及与ORF 1、2和4相关的序列是多拷贝的，并且在 B分离株之间的组织和基因组位置可变。 burgdorferi sensu lato复合体。这些分析为 B的质粒组分的高度可变的组织提供了额外的证据。伯氏疏螺旋体

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