Yang Ruifu, Lai Bipeng, Liao Kang, Liu Baomo, Huang Lixia, Li Shaoli, Gu Jincui, Lin Ziying, Chen Yili, Wang Shuaishuai, Qiu Yanli, Deng Jiating, Chen Simin, Zhuo Chao, Zhou Yanbin
Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Department of Clinical Laboratory, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, China.
Front Microbiol. 2022 Apr 25;13:867770. doi: 10.3389/fmicb.2022.867770. eCollection 2022.
, a strictly aerobic, non-lactose fermented Gram-negative bacteria, is one of the important pathogens of nosocomial infection. Major facilitator superfamily (MFS) transporter membrane proteins are a class of proteins that widely exists in microbial genomes and have been revealed to be related to biofilm formation in a variety of microorganisms. However, as one of the MFS transporter membrane proteins, little is known about the role of BIT33_RS14560 in . To explore the effects of BIT33_RS14560 on biofilm formation of , the biofilm formation abilities of 62 isolates were firstly investigated and compared with their transcript levels of . Then, this specific gene was over-expressed in a standard strain (ATCC 19606) and two isolates of extensively drug-resistant (XDR-Ab). Bacterial virulence was observed using a infection model. High-throughput transcriptome sequencing (RNA seq) was performed on ATCC 19606 over-expressed strain and its corresponding empty plasmid control strain. correlation analysis indicated a significant negative correlation (, ) between the △CT levels of BIT33_RS1456 and biofilm grading of isolates. The amount of biofilm was relatively high within 12-48 h. Regardless of standard or clinical strains; the biofilm biomass in the overexpression group was significantly higher than that in the control group (). Kaplan-Meier survival curve analysis showed that the mortality of was significantly higher when infected with the overexpression strain (). RNA-Seq showed that the mRNA expression levels of three genes annotated as OprD family outer membrane porin, glycosyltransferase family 39 protein, and glycosyltransferase family 2 protein, which were related to bacterial adhesion, biofilm formation, and virulence, were significantly upregulated when was over-expressed. Our findings provided new insights in identifying potential drug targets for the inhibition of biofilm formation. We also developed a practical method to construct an over-expressed vector that can stably replicate in XDR-Ab isolates.
[细菌名称]是一种严格需氧、不发酵乳糖的革兰氏阴性菌，是医院感染的重要病原菌之一。主要易化子超家族（MFS）转运蛋白膜蛋白是一类广泛存在于微生物基因组中的蛋白质，已被揭示与多种微生物的生物膜形成有关。然而，作为MFS转运蛋白膜蛋白之一，关于BIT33_RS14560在[细菌名称]中的作用知之甚少。为了探究BIT33_RS14560对[细菌名称]生物膜形成的影响，首先研究了62株分离株的生物膜形成能力，并将其与[细菌名称]的转录水平进行比较。然后，在标准[细菌名称]菌株（ATCC 19606）和两株广泛耐药[细菌名称]（XDR-Ab）分离株中过表达该特定基因。使用[感染模型名称]感染模型观察细菌毒力。对ATCC 19606过表达菌株及其相应的空质粒对照菌株进行高通量转录组测序（RNA seq）。相关性分析表明，BIT33_RS1456的△CT水平与[细菌名称]分离株的生物膜分级之间存在显著负相关（，）。在12至48小时内，[细菌名称]生物膜的量相对较高。无论标准菌株还是临床菌株，[细菌名称]过表达组的生物膜生物量均显著高于对照组（）。Kaplan-Meier生存曲线分析表明，感染[细菌名称]过表达菌株时，[宿主名称]的死亡率显著更高（）。RNA-Seq显示，当[细菌名称]过表达时，三个注释为OprD家族外膜孔蛋白、糖基转移酶家族39蛋白和糖基转移酶家族2蛋白的基因的mRNA表达水平显著上调，这些基因与细菌粘附、生物膜形成和毒力有关。我们的研究结果为鉴定抑制生物膜形成的潜在药物靶点提供了新的见解。我们还开发了一种实用的方法来构建一种可以在XDR-Ab分离株中稳定复制的过表达载体。
