Abstract
An artificial Mu transposition complex technique was utilized to study a cluster of genes required for swimming motility mediated by flagellum in Pseudomonas aeruginosa. Optimum electroporation conditions, including growth stages of the recipient cells and electroshock voltages, were investigated for a clinical isolate of Pseudomonas aeruginosa PA68 with artificial Mu transposition complexes. The highest electroporation efficiency was obtained at 3.66 × 104 CFU/μg DNA and a mutant library was established. Three swimming motility deficient mutants were screened from the library. Southern blot analysis shows the insertion of artificial Mini-Mu transposon into genomic DNA in a single copy. Gene cloning and sequencing of the region flanking the insertion revealed that Mini-Mu integrated into gene uvrD, zwf and phzF1, respectively. Mu transposase complexes technique is a new powerful strategy to study the function of bacterial genome. This is the first time it has been utilized to study the genes involved in the swimming motility in Pseudomonas aeruginosa.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Costerton J W, Stewart P S, Greenberg E P (1999). Bacterial biofilms: a common cause of persistent infections. Science, 284, 1318–1322
Fleiszig S M, Arora S K, Van R, Ramphal R (2001). FlhA, a component of the flagellum assembly apparatus of Pseudomonas aeruginosa, plays a role in internalization by corneal epithelial cells. Infection and immunity, 69, 4931–4937
Haapa S, Taira S, Heikkinen E, Savilahti H (1999). An efficient and accurate integration of mini-Mu transposons in vitro: a general methodology for functional genetic analysis and molecular biology applications. Nucleic Acids Research, 27, 2777–2784
Lamberg A, Nieminen S, Qiao M, Savilahti H (2002). Efficient insertion mutagenesis strategy for bacterial genomes involving electroporation of in vitro-assembled DNA transposition complexes of bacteriophage Mu. Applied Environmental Microbiology, 68, 705–712
O’Toole G A, Kolter R (1998). Flagellar and twitching motility are necessary for Pseudomonas aeruginosa biofilm development. Molecular Microbiology, 30, 295–304.
Rashid M H, Kornberg A (2000). Inorganic polyphosphate is needed for swimming, swarming, and twitching motilities of Pseudomonas aeruginosa. Proc Natl Acad Sci USA 97, 4885–4890
Sambrook J, Fritsch E F, Maniatis T, (1989). Molecular Cloning: a Laboratory Manual. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
Semmler A B, Whitchurch C B, Leech A J, Mattick J S (2000). Identification of a novel gene, fimV, involved in twitching motility in Pseudomonas aeruginosa. Microbiology 146, 1321–1332
Simpson D A & Speert D P (2000). RpmA is required for nonopsonic phagocytosis of Pseudomonas aeruginosa. Infection and immunity 68, 2493–2502
Smith A W, Iglewski B H (1989). Transformation of Pseudomonas aeruginosa by electroporation. Nucleic Acids Research, 17, 10509
Tomich M, Herfst C A, Golden J W, Mohr C D (2002). Role of flagella in host cell invasion by Burkholderia cepacia. Infection and immunity 70, 1799–1806
Author information
Authors and Affiliations
Corresponding author
Additional information
__________
Translated from Acta Scientiarum Naturalium Universitatis Nankaiensis, 2006, 39(6), 20–25 [译自: 南开大学学报(自然科学版)]
Rights and permissions
About this article
Cite this article
Shan, Z., Qiao, M., Xu, H. et al. The study of genes involved in swimming motility of Pseudomonas aeruginosa . Front. Biol. (2008). https://doi.org/10.1007/s11515-008-0059-1
Published:
DOI: https://doi.org/10.1007/s11515-008-0059-1