Construction of a Mutant Library of Avibacterium paragallinarum Transposons and Screening and Preliminary Study of Genes Related to Biofilm Formation

Abstract:This dataset supports a study investigating the genetic determinants of biofilm formation in Avibacterium paragallinarum, the causative agent of infectious coryza in poultry. A high-density random mutant library was constructed using mini-Tn5 transposon mutagenesis, comprising 3,106 independent mutants. Phenotypic screening via crystal violet staining identified 188 mutants with significantly altered biofilm-forming capacity compared to the wild-type strain. Transposon insertion sites were mapped using thermal asymmetric interlaced PCR (TAIL-PCR) and Sanger sequencing, revealing 105 unique disrupted genes. These genes are involved in diverse biological processes, including amino acid metabolism, quorum sensing, transmembrane transport, and surface structure biosynthesis. Selected mutants were further characterized for growth, adhesion, invasion, and cytotoxicity. This dataset provides a valuable resource for functional genomics studies and the development of anti-biofilm strategies against Av. paragallinarum.Experimental Design:Mutagenesis: Triparental conjugation using E. coli S17-1λpir (pEVS170) as donor, E. coli DH5αλpir (pRK2013) as helper, and Av. paragallinarum JZIC-005 as recipient.Screening: Biofilm formation assessed by crystal violet staining in 96-well plates.Insertion Site Mapping: TAIL-PCR with nested primers and Sanger sequencing.Phenotypic Assays: Growth curves, adhesion, invasion (gentamicin protection), and cytotoxicity (LDH release) in DF-1 cells.Data Features:Number of mutants: 3,106Mutants with altered biofilm: 188 (172 enhanced, 16 reduced)Unique genes identified: 105Sequencing method: TAIL-PCR + Sanger sequencingReference genome: Av. paragallinarum C-AP4 (NCBI accession: CP113955.1)