The mariner transposon family of Himar1 has been widely used for the random mutagenesis of bacteria to generate single insertions into the chromosome. Here, a versatile toolbox of mariner transposon derivatives was generated and applied to the functional genomics investigation of fish pathogen Edwardsiella piscicida. In this study, we combined the merits of the random mutagenesis of mariner transposon and common efficient reporter marker genes or regulatory elements, mcherry, gfp, luxAB, lacZ, sacBR, and P_BAD and antibiotic resistance cassettes to construct a series of derivative transposon vectors, pMmch, pMKGR, pMCGR, pMXKGR, pMLKGR, pMSGR, and pMPR, based on the initial transposon pMar2xT7. The function and effectiveness of the modified transposons were verified by introducing them into E. piscicida EIB202. Based on the toolbox, a transposon insertion mutant library containing approximately 3.0 × 10⁵ separated mutants was constructed to explore the upstream regulators of esrB, the master regulator of the type III and type VI secretion systems (T3/T6SS) in E. piscicida. Following analysis by Con-ARTIST, ETAE_2184 (renamed as EsrR) was screened out and identified as a novel regulator mediating T3SS expression. In addition, the esrR mutants displayed critical virulence attenuation. Due to the broad-range host compatibility of mariner transposons, the newly built transposon toolbox can be broadly applied for functional genomics studies in various bacteria.

Himar1的水手转座子家族已被广泛用于细菌的随机诱变，以在染色体中产生单个插入。在这里，生成了水手转座子衍生物的多功能工具箱，并将其应用于鱼类病原体爱德华氏菌的功能基因组学研究。在这项研究中，我们结合了水手转座子随机诱变的优点和常见的有效报道基因或调控元件，mcherry，gfp，luxAB，lacZ，sacBR和P _BAD以及抗生素抗性盒，构建了一系列衍生的转座子载体，第MMCH，p MKGR，p MCGR，p MXKGR，p MLKGR，p MSGR和p MPR，基于最初的转座子p Mar2xT7。通过将修饰的转座子引入到E. piscicida EIB202中来验证其功能和有效性。在该工具箱的基础上，构建了一个包含约3.0×10 ^5个分离的突变体的转座子插入突变体文库，以探索esrB的上游调节子，escB的上游调节子，E。piscicida的III型和VI型分泌系统（T3 / T6SS）。。由Con-ARTIST分析后，筛选出ETAE_2184（重命名为EsrR）并鉴定为介导T3SS表达的新型调节剂。此外，esrR突变体显示出严重的毒力衰减。由于水手转座子具有广泛的宿主相容性，新构建的转座子工具箱可广泛用于各种细菌的功能基因组学研究。