A new type of flagellin gene in Pseudomonas putida

The Journal of General and Applied Microbiology

Urai Chaisri²  Sakol Panyim¹  Sumalee Tungpradabkul¹  Saengchan Senapin¹ 

[1] Department of Biochemistry, Faculty of Science, Mahidol University;Department of Tropical Pathology, Faculty of Tropical Medicine, Mahidol University

关键词: electron microscopy;    flagellin gene;    motility inhibition assay;    polymerase chain reaction;    Pseudomonas putida;   

DOI  :  10.2323/jgam.45.105

学科分类：微生物学和免疫学

来源: Applied Microbiology, Molecular and Cellulrar Biosciences Research Foundation

【 摘 要 】

Previously established PCR amplification and Southern hybridization procedures were developed for the isolation of the 0.8-kb flagellin gene in Pseudomonas putida. The deduced protein sequence has significant homology to the N-and C-terminal sequences of other bacterial flagellins. We propose that P. putida flagellin genes can be divided at least into three size groups: type I (2.0 kb), type II (1.4 kb), and type III (0.8 kb). Type I and type II flagellin genes have been reported. The new 0.8-kb type III gene was expressed in E. coli, and the resulting protein was purified and used to raise polyclonal antibody to study whether this small gene encodes flagellin. The antiserum reacted with purified flagellin monomers from representatives of each flagellin type, as well as proteins of the same sizes in lysates of these organisms, on Western immunoblots. This antiserum was determined to be functional in a motility inhibition assay. Similar results were obtained from antiserum directed against purified type III flagellin, indicating that a new type of flagellin gene in P. putida has been found. Preliminary electron microscopic study revealed that P. putida isolate with the smaller flagellin gene type appeared to have a thinner flagellar filament.

【 授权许可】

Unknown