Grillová Linda, Romeis Emily, Lieberman Nicole A P, Tantalo Lauren C, Xu Linda H, Molini Barbara, Trejos Aldo T, Lacey George, Goulding David, Thomson Nicholas R, Greninger Alexander L, Giacani Lorenzo
bioRxiv. 2024 May 29:2024.05.29.596454. doi: 10.1101/2024.05.29.596454.
The recently discovered methodologies to cultivate and genetically manipulate subsp. ( ) have significantly helped syphilis research, allowing the evaluation of antibiotic efficacy, performance of controlled studies to assess differential treponemal gene expression, and generation of loss-of-function mutants to evaluate the contribution of specific genetic loci to virulence. Building on this progress, we engineered the SS14 strain to express a red-shifted Green Fluorescent Protein (GFP) and Sf1Ep cells to express mCherry and blue fluorescent protein (BFP) for enhanced visualization. These new resources improve microscopy- and cell sorting-based applications for , better capturing the physical interaction between the host and pathogen, among other possibilities. Continued efforts to develop and share new tools and resources are required to help our overall knowledge of biology and syphilis pathogenesis reach that of other bacterial pathogens, including spirochetes.
By employing genetic engineering, was modified to express GFP, and Sf1Ep cells to express mCherry on the cytoplasmic membrane and BFP in the nucleus. These new resources for syphilis research will facilitate experimental designs to better define the complex interplay between and the host during infection.
最近发现的用于培养和基因操作梅毒螺旋体亚种（苍白亚种）的方法极大地推动了梅毒研究，使得抗生素疗效得以评估、开展对照研究以评估梅毒螺旋体基因表达差异、生成功能丧失突变体以评估特定基因位点对毒力的作用。基于这一进展，我们对SS14菌株进行改造使其表达红移型绿色荧光蛋白（GFP），对Sf1Ep细胞进行改造使其表达mCherry和蓝色荧光蛋白（BFP）以增强可视化效果。这些新资源改善了基于显微镜检查和细胞分选的梅毒螺旋体应用，能更好地捕捉宿主与病原体之间的物理相互作用以及其他可能性。需要持续努力开发和共享新工具及资源，以帮助我们对梅毒螺旋体生物学和梅毒发病机制的整体认识达到其他细菌病原体（包括螺旋体）的水平。
通过基因工程，梅毒螺旋体被改造为表达GFP，Sf1Ep细胞被改造为在细胞质膜上表达mCherry且在细胞核中表达BFP。这些用于梅毒研究的新资源将有助于实验设计，以便更好地界定感染期间梅毒螺旋体与宿主之间复杂的相互作用。
