杜玲, 张加雪, 袁灵月, 谌丽妃, 李迎丽, 邱景富. 肺炎克雷伯菌铁摄取调节蛋白(Fur)的生物学功能及对fyuA的转录调控[J]. 云南大学学报(自然科学版), 2022, 44(1): 195-202. doi: 10.7540/j.ynu.20210024
引用本文: 杜玲, 张加雪, 袁灵月, 谌丽妃, 李迎丽, 邱景富. 肺炎克雷伯菌铁摄取调节蛋白(Fur)的生物学功能及对fyuA的转录调控[J]. 云南大学学报(自然科学版), 2022, 44(1): 195-202. doi: 10.7540/j.ynu.20210024
DU Ling, ZHANG Jia-xue, YUAN Ling-yue, SHEN Li-fei, LI Ying-li, QIU Jing-fu. Biological function of ferric uptake regulator (Fur) in Klebsiella pneumonia and the regulatory effect on fyuA[J]. Journal of Yunnan University: Natural Sciences Edition, 2022, 44(1): 195-202. DOI: 10.7540/j.ynu.20210024
Citation: DU Ling, ZHANG Jia-xue, YUAN Ling-yue, SHEN Li-fei, LI Ying-li, QIU Jing-fu. Biological function of ferric uptake regulator (Fur) in Klebsiella pneumonia and the regulatory effect on fyuA[J]. Journal of Yunnan University: Natural Sciences Edition, 2022, 44(1): 195-202. DOI: 10.7540/j.ynu.20210024

肺炎克雷伯菌铁摄取调节蛋白(Fur)的生物学功能及对fyuA的转录调控

Biological function of ferric uptake regulator (Fur) in Klebsiella pneumonia and the regulatory effect on fyuA

  • 摘要: 铁摄取调节蛋白(ferric uptake regulator, Fur)是细菌重要的调控因子,研究旨在探究肺炎克雷伯菌Fur对铁摄入及生物膜形成的影响,分析Fur对可能的靶基因fyuA(耶尔森菌素的受体)的调控作用. 首先,采用同源重组技术构建突变株Δfur及回补株C-Δfur. 然后,采用生长曲线、CAS检测和结晶紫染色对野生株、突变株及回补株的铁摄入及生物膜形成表型进行测定. 最后,通过RT-qPCR及GFP报告基因融合实验明确Fur对fyuA的调控. 结果显示,成功构建突变株及回补株;生长曲线分析表明fur基因影响细菌生长速度;fur的缺失使铁载体的生成量增加,生物膜的形成减少;铁充足条件下,Fur负调控fyuA基因的表达. 研究通过构建fur基因敲除株,初步分析了肺炎克雷伯菌铁摄取调节蛋白Fur的功能,明确了Fur对fyuA的转录调控机制.

     

    Abstract: The ferric uptake regulator (Fur) is an important regulation factor in bacteria. To explore the effect of fur gene on iron uptake and biofilm formation ability of Klebsiella pneumonia, and analyze the regulatory effect of Fur on the possible target gene fyuA (receptor for Yersinia), the unmarked fur-deletion mutant and complementation strain C-Δfur were constructed through homologous recombination. Phenotype of wild strains, mutant strains and replenishment strains was determined by growth curve, CAS detection and crystal violet staining, and the transcription regulation mechanism of Fur on fyuART-q was clarified by PCR and GFP fusion assay. The results showed that fur affected the growth rate of bacteria; Deletion of the fur gene increased the production of siderophores and reduced the biofilm formation; The fyuA gene was negatively regulated by Fur under iron sufficient conditions.

     

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