摘要
【目的】探索EnvZ/OmpR双组分调控系统的效应蛋白OmpR对粘质沙雷氏菌FZSF02灵菌红素合成及其他生物学特性的影响。【方法】同源重组法构建ompR敲除菌株,琼脂平板产色试验和qPCR检测OmpR对菌株灵菌红素合成的影响,结晶紫染色法和琼脂平板法等研究基因敲除菌株生物被膜形成能力,运动性和对不同环境胁迫因素的耐受性。【结果】序列分析显示OmpR为序列高度保守的蛋白。PCR验证证明ompR基因敲除成功;与野生型菌株相比ΔompR失去灵菌红素合成能力,qPCR试验显示灵菌红素合成基因簇中3个关键基因pigA、pigF和pigN转录水平分别降低为野生型菌株的3.8%,2.0%和2.1%;ΔompR菌株生物被膜生成能力较野生型降低37.5%(37℃)和15.1%(27℃);OmpR对该菌的生长能力、运动能力和响应环境胁迫的能力无明显影响。【结论】ompR为一新报道的特异性调控粘质沙雷氏菌灵菌红素合成的基因,且该基因对该菌生物被膜的形成有重要影响。
【Objective】Biological functions of the regulatory protein ompR in the two component EnvZ/OmpR system,including prodigiosin-producing ability and other biological characteristics,of Serratia marcescens FZSF02 were studied.【Methods】Homologous recombination was used to construct ompR-knockout S.marcescens FZSF02.Effect of OmpR on the prodigiosin-producing ability was examined by LB agar plate incubation and qPCR.Methods of crystal violet staining,agar plate incubation,and others were applied to determine the biofilm-forming,mobility,and stress adaptation abilities of the transcription factor protein under various environmental stresses.【Results】OmpR was a protein with high conserved amino acid sequences.An ompR-deleted strain,FZSF02ΔompR,was successfully obtained by homologous recombination and confirmed by PCR.As a result,FZSF02ΔompR lost prodigiosin-producing ability that possessed by the wild strain.The transcriptional levels of pigA,pigF,and pigN of the prodigiosin biosynthesis gene cluster in FZSF02ΔompR were respectively 3.8%,2.0%and 2.1%of the wild type strain.The biofilm formation of FZSF02ΔompR declined 37.5%(at 37℃)and 15.1%(at 27℃)from its wild counterpart.On the other hand,OmpR exhibited no significant effect on the growth,mobility,or response to the environmental stress.【Conclusion】OmpR was a newly reported gene that specifically regulated the prodigiosin biosynthesis in S.marcescens.It also significantly affected the biofilm formation but not on the growth,mobility,or stress response.
作者
贾宪波
刘方晨
赵恪
林俊杰
方宇
陈济琛
JIA Xianbo;LIU Fangchen;ZHAO Ke;LIN Junjie;FANG Yu;CHEN Jichen(Institute of Soil and Fertilizer,Fujian Academy of Agricultural Sciences,Fuzhou,Fujian 350003,China;College of Resources and Environment,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350001,China;College of Life Sciences,Fujian Agriculture and Forestry University,Fuzhou,Fujian 350001,China)
出处
《福建农业学报》
CAS
CSCD
北大核心
2021年第12期1491-1498,共8页
Fujian Journal of Agricultural Sciences
基金
国家自然科学基金青年基金项目(31800068)
福建省农业科学院科技创新团队建设项目(CXTD2021002-3)。