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冯宇晴, 张倩倩, 章晋勇, 张兰梨, 马丽娜, 冯东岳, 吴振兵, 郝静文, 王姝懿, 李爱华. AmoA、AmoE和AmoF蛋白在嗜水气单胞菌铁载体合成中的作用研究[J]. 水生生物学报, 2019, 43(3): 486-493. DOI: 10.7541/2019.060
引用本文: 冯宇晴, 张倩倩, 章晋勇, 张兰梨, 马丽娜, 冯东岳, 吴振兵, 郝静文, 王姝懿, 李爱华. AmoA、AmoE和AmoF蛋白在嗜水气单胞菌铁载体合成中的作用研究[J]. 水生生物学报, 2019, 43(3): 486-493. DOI: 10.7541/2019.060
FENG Yu-Qing, ZHANG Qian-Qian, ZHANG Jin-Yong, ZHANG Lan-Li, MA Li-Na, FENG Dong-Yue, WU Zhen-Bing, HAO Jing-Wen, WANG Shu-Yi, LI Ai-Hua. ROLE OF AMOA, AMOE, AND AMOF PROTEINS FOR SIDEROPHORE SYNTHESIS OF AEROMONAS HYDROPHILA[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(3): 486-493. DOI: 10.7541/2019.060
Citation: FENG Yu-Qing, ZHANG Qian-Qian, ZHANG Jin-Yong, ZHANG Lan-Li, MA Li-Na, FENG Dong-Yue, WU Zhen-Bing, HAO Jing-Wen, WANG Shu-Yi, LI Ai-Hua. ROLE OF AMOA, AMOE, AND AMOF PROTEINS FOR SIDEROPHORE SYNTHESIS OF AEROMONAS HYDROPHILA[J]. ACTA HYDROBIOLOGICA SINICA, 2019, 43(3): 486-493. DOI: 10.7541/2019.060

AmoA、AmoE和AmoF蛋白在嗜水气单胞菌铁载体合成中的作用研究

ROLE OF AMOA, AMOE, AND AMOF PROTEINS FOR SIDEROPHORE SYNTHESIS OF AEROMONAS HYDROPHILA

  • 摘要: 铁载体被认为是嗜水气单胞菌的毒力因子之一, 其由amoCEBFAGH七个基因编码, AmoCGH在前人的研究中已证实参与铁载体的合成。RT-PCR实验表明amoAEF基因的表达受到铁的调控。为进一步探究amoAEF基因的功能, 利用融合PCR和基因同源重组原理, 以自杀性质粒PRE112为载体构建基因缺失株ΔamoA、ΔamoE和ΔamoF。通过CAS平板检测实验以及arnow实验来检测野生株WT与各基因缺失突变株铁载体的合成情况, 并比较野生株与各缺失株在低铁培养基中的生长差异。结果显示, 成功构建了基因缺失株ΔamoA、ΔamoE和ΔamoF; 在富铁条件下, 基因缺失株ΔamoA、ΔamoE和ΔamoF的生长与野生株无显著性差异, 但在低铁条件下, 基因缺失株ΔamoA、ΔamoE和ΔamoF的生长能力、铁载体合成能力显著低于野生株。可见, amoAamoEamoF基因是嗜水气单胞菌铁载体合成的关键基因, 其缺失会导致细菌在低铁环境中的生长受到抑制。

     

    Abstract: Siderophore is considered as one of the virulence factors of Aeromonas hydrophila and is encoded by seven genes of amoCEBFAGH. AmoCGH have been confirmed to be involved in the synthesis of siderophore in previous studies. RT-PCR assay indicated that the expression of amoAEF genes were regulated by iron. In order to further explore the function of genes amoAEF, in this study, we used the suicide plasmid PRE112 to construct gene deletion mutant strains ΔamoA, ΔamoE and ΔamoF based on the infusion PCR and gene homologous recombination principle. The detection of the siderophore synthesis in the wild strains and mutant strains were performed using CAS plate test and arnow test. The results showed that Aeromonas hydrophila ΔamoA, ΔamoE and ΔamoF were successfully constructed. The growth of wild strain, ΔamoA, ΔamoE and ΔamoF had no significant difference in iron-riched medium, while in iron-depleted medium, the growth and siderophore synthesis capacity of ΔamoA, ΔamoE and ΔamoF were significantly lower than that of the wild strain. The results suggested that amoA, amoE and amoF are the critical synthesis genes of siderophore, and the growth of the deletion mutants were inhibited under low iron conditions.

     

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