Bueno Danilo, Pedrolli Danielle B, Martins Paula M M, Bocchini Daniela A, Moraes Karen C M, Facincani Agda P, Ferro Jesus A, Varani Alessandro M, Pena Michelle M, Ferreira Henrique
Department of General and Applied Biology, Biosciences Institute, São Paulo State University (UNESP), Rio Claro SP 13506-900, Brazil.
Department of Biological Sciences, School of Pharmaceutical Sciences, São Paulo State University (UNESP), Araraquara SP 14800-903, Brazil.
Microorganisms. 2021 Feb 6;9(2):329. doi: 10.3390/microorganisms9020329.
subsp. () is the causal agent of Asiatic Citrus Canker (ACC), a disease that affects citrus. ACC has no cure, and growers must rely on special agricultural practices to prevent bacterial spreading. Understanding basic biology is essential to foresee potential genetic targets to control ACC. Traditionally, microbial genetics use gene deletion/disruption to investigate gene function. However, essential genes are difficult to study this way. Techniques based on small-RNAs and antisense-RNAs are powerful for gene characterization, but not yet fully explored in prokaryotes. One alternative is riboswitches, which derive from bacteria, and can control transcription/translation. Riboswitches are non-coding RNAs able to modulate gene expression in the presence of specific ligands. Here we demonstrate that the riboswitch theo/ decreases expression in in a platform responsive to theophylline. By monitoring cell respiration, we showed that higher concentrations of the ligand interfered with bacterial viability. Therefore, we determined the safe dose of theophylline to be used with . Finally, in downstream investigations of transcription modulation, we show evidence for the fact that ParB is stable, remains functional throughout the cell cycle, and is inherited by the daughter cells upon cell division.
亚种（）是亚洲柑橘溃疡病（ACC）的病原体，这种疾病会影响柑橘类植物。ACC无法治愈，种植者必须依靠特殊的农业措施来防止细菌传播。了解其基本生物学特性对于预测控制ACC的潜在基因靶点至关重要。传统上，微生物遗传学利用基因缺失/破坏来研究基因功能。然而，必需基因很难通过这种方式进行研究。基于小RNA和反义RNA的技术在基因表征方面很强大，但在原核生物中尚未得到充分探索。一种替代方法是核糖开关，它源自细菌，能够控制转录/翻译过程。核糖开关是一种非编码RNA，能够在特定配体存在的情况下调节基因表达。在这里，我们证明了核糖开关theo/在对茶碱有反应的平台中降低了在中的表达。通过监测细胞呼吸，我们发现较高浓度的配体干扰了细菌的生存能力。因此，我们确定了与一起使用的茶碱的安全剂量。最后，在对转录调控的下游研究中，我们证明了ParB是稳定的，在整个细胞周期中都保持功能，并在细胞分裂时由子细胞继承。
