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首页> 外文期刊>Archives of microbiology >Isolation of transposon mutants and characterization of genes involved in biofilm formation by Pseudomonas fluorescens TC222.
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Isolation of transposon mutants and characterization of genes involved in biofilm formation by Pseudomonas fluorescens TC222.

机译：转座子突变体的分离和荧光假单胞菌TC222参与生物膜形成的基因的表征。

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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.
机译：通常发现生物膜形成突变体具有缺陷或改变的运动性。利用运动型来鉴定荧光假单胞菌生物膜形成突变体。从荧光假单胞菌分离物TC222中获得了十四个活力突变体，并进一步研究了八个稳定突变体。与亲本相比，八个转座子插入突变体显示形成生物膜的能力发生了改变。克隆并测序了来自这些突变体的五个Tn5插入的基因。遗传分析表明，两个插入位于影响多个细胞表面特征的基因中，包括脂多糖（rfbD）和极地鞭毛（fliR）。首次报道了编码假定的Mig-14家族蛋白（epsB），可能的噬菌体信号肽蛋白（bspA）和可溶性吡啶核苷酸转氢酶（pyrA）的三个基因与生物膜形成有关。 rfbD和epsB基因的互补实验证明，可以恢复相应突变体的生物膜形成。进一步的半定量逆转录-PCR分析表明，rfbD和epsB均可在互补菌株中表达其转录本，其表达水平远高于野生型菌株。无法检测到两个基因在其突变体中的转录本。

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