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Microbiology 150 (2004), 2653-2661; DOI  10.1099/mic.0.27131-0
© 2004 Society for General Microbiology

Identification of two new genes involved in twitching motility in Pseudomonas aeruginosa

Zhiying Shan1,2, Haijin Xu1, Xingqi Shi1, Yan Yu1, Hongming Yao1, Xiuming Zhang1, Yanling Bai1, Caichang Gao1, Per E. J. Saris3 and Mingqiang Qiao1,3

1 Institute for Molecular Biology, Nankai University, Tianjin, People's Republic of China
2 The Second Hospital of Tianjin Medical University, Tianjin, People's Republic of China
3 Department of Applied Chemistry and Microbiology, Fin-00014 University of Helsinki, Finland

Correspondence
Mingqianq Qiao
mingqiangqiao{at}yahoo.com.cn

Mu transposition complexes were used for transposon mutagenesis of Pseudomonas aeruginosa strain PA68. Mu DNA transposition complexes were assembled with MuA transposase and an artificial mini-Mu transposon in vitro, and introduced into Pseudomonas aeruginosa by electroporation. Eight mutants deficient in twitching motility were isolated. Southern blotting confirmed that the insertions had occurred as single events. DNA sequencing of the region flanking the insertion in the twitching-motility mutants revealed that the mini-Mu transposon had inserted into six different genes, PAO171, PA1822, PAO413, PA4959, PA4551 and PA5040. Four of these have previously been proven to be needed for twitching motility, whereas the PA1822 and PA0171 genes have not previously been shown to be required for twitching motility. The twitching-motility defect in the PA1822 mutant was partially complemented by providing the PA1822 gene in trans, and the defect in the PA0171 mutant was fully complemented when PA0171 was provided. A PA0171 mutant and a PA1822 mutant were constructed by gene replacement in the P. aeruginosa PAO1 strain. These mutants were deficient in twitching motility, showing that both the PA1822 and the PA0171 gene are involved in twitching motility.

The GenBank/EMBL/DDBJ accession numbers for the 16S rRNA gene, PA0171 and PA1822 sequences reported in this paper are AY521230, AY502957 and AY502958, respectively.






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Copyright © 2004 Society for General Microbiology.