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外膜蛋白A (OmpA)在细菌的致病中表现出多种功能,包括对宿主细胞的粘附、侵袭等,是介导病原菌与宿主相互作用的关健因子。本研究以维氏单胞菌C4基因组DNA为模板,扩增330 bp和442 bp OmpA的上下游同源臂片段,成功构建自杀重组质粒pRE112-ΔOmpA;将重组质粒转化至E. coli WM3064,再双亲接合至维氏气单胞菌中,基于同源重组的原理在8%蔗糖板上成功筛选得到维氏气单胞菌C4 OmpA基因敲除菌ΔOmpA。生长特性测定结果表明敲除株ΔOmpA较野生菌株WT生长更为缓慢,本研究结果为进一步研究OmpA基因在维氏单胞菌中的功能提供了科学依据。
Abstract：The outer membrane protein A is a key factor mediating the interaction between pathogens and host,and it exhibits multiple functions in the pathogenesis of bacteria, including adhesion and invasion to host cells. In this study, the 330 bp upstream and 442 bp downstream of the OmpA gene were amplified from genomic DNA of A. veronii C4, and inserted to the backbone to construct the pRE112-ΔOmpA plasmid successfully. The recombinant construction was transformed into E. coli WM3064 and then mobilized into A. veronii C4 via biparental mating,and the deletion mutants were selected on LB medium containing 8% sucrose. Growth characteristics showed thatΔOmpA grows more slowly than WT. The results of this study provide scientific basis for further research on the function of OmpA gene in A eromonas veronii C4.
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基本信息:
DOI：10.13417/j.gab.039.003505
中图分类号:Q93
引用信息:
[1]许琪,马香,李宏,等.维氏气单胞菌C4外膜蛋白A基因敲除株的构建及其生长特性[J].基因组学与应用生物学,2020,39(08):3505-3509.DOI:10.13417/j.gab.039.003505.
基金信息:
海南省自然科学基金(317015);; 海南省重点研发计划(ZDYF2017020);; 国家自然科学基金项目(31360261;31560021; 31772887)共同资助
2019-03-12
2019-03-12
2019-03-12