Gene Expression Platform for Synthetic Biology in the Human Pathogen Streptococcus pneumoniae
Gene Expression Platform for Synthetic Biology in the Human Pathogen Streptococcus pneumoniae
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DOI：
10.1021/sb500229s
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发表时间：
2015-03-01
影响因子：
4.7
通讯作者：
Veening, Jan-Willem
中科院分区：
文献类型：
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作者：
Sorg, Robin A.;Kuipers, Oscar P.;Veening, Jan-Willem
人类病原体肺炎链球菌（肺炎球菌）是一种细菌，其成功归功于细胞和群体水平上的复杂基因表达调控模式。毒力因子的表达使大部分无危害的存在，与宿主和生态位竞争对手的平衡。在特定情况下，这种表达的变化可能导致更具侵略性的行为和病原体入侵形式的逆转。这些触发条件非常难以研究，因为环境线索通常是未知的或几乎不可能在宿主外部（体外）模拟。研究表达模式的另一种方法是在合成生物学方法中发现的，即用正交成分模拟观察到的行为来重建调控网络。在这里，我们创建了一个适用于肺炎链球菌合成生物学方法的遗传平台，并表征了一组标准化的启动子和报告基因。我们表明，我们的系统允许快速和容易的克隆与BglBrick系统和可靠和强大的基因表达后，整合到肺炎链球菌基因组中实现。此外，克隆系统被扩展以允许核糖体结合位点、肽标签和融合蛋白的直接基于接头的组装，并且我们将这种新的普遍适用的标准称为“BglFusion”。本研究中描述的基因表达平台和方法为在肺炎链球菌中采用合成生物学方法铺平了道路。
The human pathogen Streptococcus pneumoniae (pneumococcus) is a bacterium that owes its success to complex gene expression regulation patterns on both the cellular and the population level. Expression of virulence factors enables a mostly hazard-free presence of the commensal, in balance with the host and niche competitors. Under specific circumstances, changes in this expression can result in a more aggressive behavior and the reversion to the invasive form as pathogen. These triggering conditions are very difficult to study due to the fact that environmental cues are often unknown or barely possible to simulate outside the host (in vitro). An alternative way of investigating expression patterns is found in synthetic biology approaches of reconstructing regulatory networks that mimic an observed behavior with orthogonal components. Here, we created a genetic platform suitable for synthetic biology approaches in Streptococcus pneumoniae and characterized a set of standardized promoters and reporters. We show that our system allows for fast and easy cloning with the BglBrick system and that reliable and robust gene expression after integration into the Streptococcus pneumoniae genome is achieved. In addition, the cloning system was extended to allow for direct linker-based assembly of ribosome binding sites, peptide tags, and fusion proteins, and we called this new generally applicable standard "BglFusion". The gene expression platform and the methods described in this study pave the way for employing synthetic biology approaches in Streptococcus pneumoniae.