Use of recombinase gene fusions to identify Vibrio cholerae genes induced during infection
Use of recombinase gene fusions to identify Vibrio cholerae genes induced during infection
复制标题
DOI：
10.1111/j.1365-2958.1995.mmi_18040671.x
复制
发表时间：
1995-11-01
影响因子：
3.6
通讯作者：
Mekalanos, JJ
中科院分区：
文献类型：
--
作者：
Camilli, A;Mekalanos, JJ
对宿主-寄生虫相互作用的完整理解必须包括在感染过程中双方表达的基因产物的鉴定和表征。我们已经开发了一种新的筛选方法来鉴定在宿主动物感染过程中转录诱导的细菌基因，该方法基于对携带tnpR操纵子融合的菌株的预先选择（编码解离酶，一种位点特异性DNA重组酶），然后筛选这些菌株的一个子集，其随后在宿主环境中表达解离酶，后一个子集被认为是删除了溶解酶特异性报告基因构建体的重组体。霍乱弧菌的13个转录单位被鉴定为在霍乱的幼鼠模型中感染期间被诱导。其中五个被预测为编码具有不同功能的代谢，生物合成和运动的多肽;一个编码分泌脂肪酶;两个似乎是反义基因参与运动;和五个被预测为编码未知功能的多肽。三个转录本被证明是所需的完整的毒力在婴儿小鼠中，通过竞争实验进行评估。
A complete understanding of host-parasite interactions must necessarily include the identification and characterization of gene products expressed by both parties during the infectious process. We have developed a new screen to identify bacterial genes that are transcriptionally induced during infection of a host animal, The method is based on pre-selection of strains carrying tnpR operon fusions (encoding resolvase, a site-specific DNA recombinase) which are not expressed in vitro, followed by screening for a subset of these strains that subsequently express resolvase within the host environment, The latter subset was recognized as recombinants that had deleted a resolvase-specific reporter construct. Thirteen transcription units of Vibrio cholerae were identified that were induced during infection in an infant mouse model of cholera. Five of these were predicted to encode polypeptides with diverse functions in metabolism, biosynthesis and motility; one encoded a secreted lipase; two appear to be antisense to genes involved in motility; and five are predicted to encode polypeptides of unknown function. Three of the transcripts were shown to be required for full virulence in infant mice, as assessed by competition experiments.