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【目的】探究kdpD基因对溶藻弧菌生物学特性的影响。【方法】采用Overlap PCR和同源重组技术,结合正负向筛选,构建kdpD基因无标记基因框内敲除突变株,比较kdpD突变株和野生株HY9901在生长速率、胞外蛋白酶活性以及毒力等方面的差异。【结果】成功构建溶藻弧菌kdpD基因敲除突变株。体外实验表明,kdpD的缺失对溶藻弧菌的生长曲线和胞外蛋白酶活性的影响不明显,但是突变株的泳动能力和生物被膜形成能力出现下降。斑马鱼致病性试验结果显示,突变株毒力下降了8.84倍。【结论】kdpD基因参与调控溶藻弧菌的泳动能力、被膜形成和毒力,但不影响生长速率和胞外蛋白酶活性。
【Objective】 To investigate the effect of kdpD gene on the biological characteristics of Vibrio alginolyticus. 【Method】 By using Overlap PCR and homologous recombination, positive and negative screening was carried out to construct knockout mutants with unlabeled kdpD gene. The growth rate, extracellular protease activity and virulence of kdpD mutant and wild strain HY9901 were compared. And so on the difference. 【Result】 The kdpD knockout mutant of V. alginolyticus was successfully constructed. In vitro experiments showed that the deletion of kdpD had no significant effect on the growth curve and extracellular protease activity of V. alginolyticus, however, the ability of migration and biofilm formation of mutant strains decreased. The results of zebrafish pathogenicity test showed that the virulence of mutants decreased by 8.84 times. 【Conclusion】 The kdpD gene is involved in the regulation of the swimming ability, membrane formation and virulence of V. alginolyticus, but not affecting the growth rate and extracellular protease activity.