The incidence of Vibrio alginolyticus infections has increased in recent years due to the influence of climate change and rising sea temperature. Vibrio virulence regulatory RNA 1 (Vvrr1) is a newly found noncoding RNA (ncRNA) predicted to be closely related to the adhesion ability of V. alginolyticus based on the previous RNA-seq. In this study, the target genes of Vvrr1 were fully screened and verified by constructing Vvrr1-overexpressing strains and using the proteome sequencing technology. Pyruvate kinase I (pykF) gene was predicted to be a chief target gene of Vvrr1 involved in virulence regulation. The adhesion ability, biofilm formation and virulence were significantly reduced in the Vvrr1-overexpressing and the pykF-silenced strain compared with the wild strains. Similar to the overexpression of Vvrr1, the silencing of pykF also reduced the expression level of virulence genes, such as ndk, eno, sdhB, glpF, and cysH. Meanwhile, by constructing the "pykF-GFP" fusion expression plasmid and using the GFP reporter gene analysis in Escherichia coli, the fluorescence intensity of the strain containing Vvrr1 whole ncRNA sequence vector was found to be significantly weakened. These indicated that Vvrr1 participated in the virulence regulation mechanism of V. alginolyticus by interacting with the virulence gene pykF.

中文翻译：

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通过比较蛋白质组分析研究新溶藻弧菌 ncRNA Vvrr1 毒力调节的机制。

近年来，由于气候变化和海水温度上升的影响，溶藻弧菌感染的发病率有所增加。弧菌毒力调节RNA 1（Vvrr1）是新发现的一种非编码RNA（ncRNA），根据之前的RNA-seq预测其与溶藻弧菌的粘附能力密切相关。本研究通过构建Vvrr1过表达菌株并利用蛋白质组测序技术对Vvrr1的靶基因进行了全面筛选和验证。丙酮酸激酶 I (pykF) 基因预计是 Vvrr1 参与毒力调节的主要靶基因。与野生菌株相比，Vvrr1过表达菌株和pykF沉默菌株的粘附能力、生物膜形成和毒力均显着降低。与Vvrr1的过度表达类似，pykF的沉默也降低了毒力基因的表达水平，例如ndk、eno、sdhB、glpF和cysH。同时，通过构建“pykF-GFP”融合表达质粒，并在大肠杆菌中使用GFP报告基因分析，发现含有Vvrr1全ncRNA序列载体的菌株的荧光强度明显减弱。这表明Vvrr1通过与毒力基因pykF相互作用参与了溶藻弧菌的毒力调控机制。