Isolation of transposon mutants and characterization of genes involved in biofilm formation by Pseudomonas fluorescens TC222

文献类型: 外文期刊

第一作者: Zhang, Jie

作者: Zhang, Jie;Song, Fuping;Fan, Liqiang;Huang, Dafang

作者机构:

关键词: pseudomonasfluorescens;Tn5 mutagenesis;biofilm formation;complementation;semi-quantitative reverse transcription-PCR analysis

期刊名称:ARCHIVES OF MICROBIOLOGY ( 影响因子:2.552; 五年影响因子:2.475 )

ISSN: 0302-8933

年卷期: 2007 年 188 卷 3 期

页码:

收录情况: SCI

摘要: Biofilm formation mutants are often found to have defective or altered motility. The motility phenotype was exploited to identify Pseudomonas fluorescens biofilm formation mutants. Fourteen motility mutants were obtained from P. fluorescens isolate TC222 and eight stable mutants were studied further. The eight transposon insertion mutants showed altered ability to form biofilm compared with the parent. Five Tn5-inserted genes from these mutants were cloned and sequenced. Genetic analysis showed that two insertions were located in genes affecting multiple cell surface characteristics, including lipopolysaccharide (rfbD) and polar flagella (fliR). Three genes encoding for a putative Mig-14 family protein (epsB), a probable bacteriophage signal peptide protein (bspA) and a soluble pyridine nucleotide transhydrogenase (pyrA) were reported for the first time to be involved in biofilm formation. Complementation experiments of rfbD and epsB genes proved that biofilm formation of the corresponding mutants could be restored. Further semi-quantitative reverse transcription-PCR analysis showed that both rfbD and epsB can express their transcripts much higher in the complemented strains than that in wild-type strains. The transcripts of both genes in their mutants could not be detected.

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