Genomic and phenotypic characterization of in vitro-generated Chlamydia trachomatis recombinants.
Genomic and phenotypic characterization of in vitro-generated Chlamydia trachomatis recombinants.
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DOI：
10.1186/1471-2180-13-142
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发表时间：
2013-06-20
期刊：
影响因子：
4.2
通讯作者：
Rockey DD
中科院分区：
文献类型：
--
作者：
Jeffrey BM;Suchland RJ;Eriksen SG;Sandoz KM;Rockey DD
前基因组和后基因组研究表明，衣原体在体外和体内积极重组，尽管这一过程的分子和细胞生物学尚不清楚。在这项研究中，我们确定了在抗生素选择下体外产生的 12 种沙眼衣原体重组体的基因组序列。这些菌株用于探索衣原体属的重组过程，包括分析候选重组热点，并将已知的沙眼衣原体体外表型与亲本表型和基因型相关联。 190 个检查的重组事件中的每一个都是同源重组的产物，并且在重组位点没有鉴定出候选靶向基序。在一个重组子代中存在单一缺失事件，导致两个 rRNA 操纵子之间 17.1 KB 的删除。没有证据表明重组偏好于染色体的任何特定区域，并且对总共 200 多个个体重组事件的分析并未为体外重组热点提供任何支持。这些研究分析了两种可测量的表型。首先，检查了在没有离心的情况下与宿主细胞的附着效率，并且该特性分离到携带多态性膜蛋白（Pmp）基因的染色体区域。其次，细胞内二次内含物的形成在重组后代中有所不同，但这并没有完全分离到染色体的特定区域。这些实验检查了沙眼衣原体的重组过程，并鉴定了可用于将重组后代的表型与基因型关联起来的工具。没有数据支持特定核苷酸序列优先用于体外重组的假设。可以通过重组分析来分离选定的表型，并且该技术可用于初步分析衣原体遗传变异与表型变异的关系。
Pre-genomic and post-genomic studies demonstrate that chlamydiae actively recombine in vitro and in vivo, although the molecular and cellular biology of this process is not well understood. In this study, we determined the genome sequence of twelve Chlamydia trachomatis recombinants that were generated in vitro under antibiotic selection. These strains were used to explore the process of recombination in Chlamydia spp., including analysis of candidate recombination hotspots, and to correlate known C. trachomatis in vitro phenotypes with parental phenotypes and genotypes. Each of the 190 examined recombination events was the product of homologous recombination, and no candidate targeting motifs were identified at recombination sites. There was a single deletion event in one recombinant progeny that resulted in the removal of 17.1 kilobases between two rRNA operons. There was no evidence for preference for any specific region of the chromosome for recombination, and analyses of a total of over 200 individual recombination events do not provide any support for recombination hotspots in vitro. Two measurable phenotypes were analyzed in these studies. First, the efficiency of attachment to host cells in the absence of centrifugation was examined, and this property segregated to regions of the chromosome that carry the polymorphic membrane protein (Pmp) genes. Second, the formation of secondary inclusions within cells varied among recombinant progeny, but this did not cleanly segregate to specific regions of the chromosome. These experiments examined the process of recombination in C. trachomatis and identified tools that can be used to associate phenotype with genotype in recombinant progeny. There were no data supporting the hypothesis that particular nucleotide sequences are preferentially used for recombination in vitro. Selected phenotypes can be segregated by analysis of recombination, and this technology may be useful in preliminary analysis of the relationship of genetic variation to phenotypic variation in the chlamydiae.
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影响因子：
3.6
作者：
Fields, KA;Mead, DJ;Hackstadt, T
通讯作者：
Hackstadt, T
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影响因子：
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通讯作者：
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通讯作者：
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作者：
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通讯作者：
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影响因子：
4.8
作者：
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通讯作者：
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