生物学杂志

生物学杂志 ›› 2020, Vol. 37 ›› Issue (6): 42-.doi: 10.3969/j.issn.2095-1736.2020.06.042

• 研究报告 • 上一篇    下一篇

维氏气单胞菌acrR敲除株的构建

  

  1. 海南大学 生命科学与药学院 热带生物资源教育部重点实验室， 海口 570228
  • 出版日期:2020-12-18 发布日期:2020-12-21
  • 通讯作者: 马香，博士，副教授，研究方向为病原微生物耐药机制，E-mail：993034@hainanu.edu.cn
  • 作者简介:胡康，硕士研究生，研究方向为病原微生物，E-mail：kanghu941015@163.com
  • 基金资助:
    海南省自然科学基金青年基金(319QN161)；国家自然科学基金项目(31772887)

The construction of acrR deletion strain of Aeromonas Veronii

  1. Key Laboratory of Tropical Biological Resources of Ministry of Education, School of Life and Pharmaceutical Sciences, Hainan University, Haikou 570228, China
  • Online:2020-12-18 Published:2020-12-21

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摘要： 从维氏气单胞菌C4(A. veronii C4)基因组DNA中扩增得到acrR基因上游同源臂片段和下游同源臂片段，连接到自杀质粒pRE112中以构建敲除重组质粒pRE112-ΔacrR，并通过电转化将敲除重组质粒导入大肠杆菌WM3064中，再通过双亲接合将其转入维氏气单胞菌C4中，以筛选得到acrR基因敲除的维氏气单胞菌突变株。生长曲线测定结果显示acrR基因敲除不会影响维氏气单胞菌的生长。生物膜测定表明，敲除acrR基因的维氏气单胞菌株表现出生物膜的形成显著增加。所构建的敲除株为进一步探索AcrR在维氏气单胞菌中的功能提供了必要材料，并提示AcrR在细菌生物膜形成中的潜在调控功能。

关键词: acrR 基因, 基因敲除, 生物膜, 维氏气单胞菌

Abstract: The recombinant plasmid pRE112-ΔacrR was constructed by amplifying the upstream and downstream homologous arms of acrR gene from the genomic DNA of Aeromonas veronii C4 and ligating them into the suicide plasmid pRE112. Subsequently the recombinant plasmid was transformed into E. coli WM3064, and mobilized into A. veronii to select the acrR gene knockout strain. Biofilm assay and growth curve analysis showed that knockout of acrR gene enhanced the biofilm formation but did not affect the growth of A. veronii. This study not only provided a necessary research material for the further exploration of the functions of AcrR in A. veronii, but also shed a light on the potential roles of AcrR in biofilm formation.

Key words: acrR gene, gene knockout, biofilm, Aeromonas veronii

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