Contribution of motA gene in pathogenesis-associated chemotaxis and colonization of Campylobacter jejuni
10.3760/cma.j.issn.0254-5101.2010.03.001
- VernacularTitle:motA基因在空肠弯曲菌致病性相关趋化和定植中的作用
- Author:
Ping RUAN
;
Aihua SUN
;
Xin ZHAO
;
Jie YAN
- Publication Type:Journal Article
- Keywords:
Campylobacter jejuni;
motA gene;
Flagelar motility;
Chemotaxis;
Colonization
- From:
Chinese Journal of Microbiology and Immunology
2010;30(3):195-200
- CountryChina
- Language:Chinese
-
Abstract:
Objective To determine the role of flagellar motor protein MotA in the pathogenesisassociated chemotaxis and colonization of Campylobacter jejuni. Methods The motA gene as well as Kan~r gene and plus-motA gene segments for motA gene knock-out were amplified by PCR and the target amplification fragments were sequenced after cloning. A suicide plasmid(pBlueskrit-Ⅱ-SK~(motA-kan)) and a motA gene knock-out mutant (motA~-) were constructed based on homologious recombination. By using semisolid plate migration test, hard agar plus (HAP)-based chemotactic test towards sodium deoxycholate (SDC) in vitro, and jejunal colonization test in BALB/c-ByJ mice were performed to determine the differences of flagellar motility, chemotaxis towards SDC and colonization in murine jejunum between motA~- mutant and wild-type strain. Results The nucleotide and amino acid sequences of the cloned motA gene were 100% identical to the reported corresponding sequences. The results of PCR, sequencing and continuous passage culture in antibiotics-contained medium demonstrated that both suicide plasmid and motA~- mutant were successfully generated. The diameters of clonies on semisolid plate and 0.2 mol/L SDC-induced chemotactic tings in HAP as well as the bacterial numbers adhering to the surface of murine jejunal mucosa and in jejunal content of motA~- mutant were significantly less than those of wild-type strain(P<0.05). Conclusion A motA gene knock-out mutant of C. jejuni was successfully constructed in this study, motA is an essential gene for flagellax motility, pathogenesis-associated chemotaxis and colonization of C. jejuni.
