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Applied and Environmental Microbiology, November 2005, p. 6633-6643, Vol. 71, No. 11
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.11.6633-6643.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Detection, Characterization, and In Vitro and In Vivo Expression of Genes Encoding S-Proteins in Lactobacillus gallinarum Strains Isolated from Chicken Crops

Karen E. Hagen ,^1,, Le Luo Guan,^1, Gerald W. Tannock,^1,2 Doug R. Korver,¹ and Gwen E. Allison^1,3,4*

Department of Agricultural, Food and Nutritional Science, University of Alberta, Alberta, Canada T6G 2P5,¹ Department of Microbiology and Immunology, University of Otago, Dunedin, New Zealand,² School of Biochemistry and Molecular Biology, Faculty of Science, The Australian National University, Canberra, ACT 0200, Australia,³ ANU Medical School, The Australian National University, Canberra, ACT 0200, Australia⁴

Received 13 February 2005/ Accepted 23 June 2005

Thirty-eight isolates of Lactobacillus gallinarum cultured from the crops of broiler chickens were screened for the presence of genes encoding S-layer proteins. All of the isolates had two S-protein genes, which were designated Lactobacillus gallinarum S-protein (lgs) genes. One gene in each isolate was either lgsA or lgsB. The Lactobacillus isolates were further characterized by pulsed-field gel electrophoresis of DNA digests, which grouped the isolates into 17 genotypes (strains). The second gene in each of eight representative strains was sequenced and shown to differ among strains (lgsC, lgsD, lgsE, lgsF, lgsG, lgsH, and lgsI). The genome of each strain thus encoded a common S-protein (encoded by either lgsA or lgsB) and a strain-specific S-protein. The extraction of cell surface proteins from cultures of the eight strains showed that each strain produced a single S-protein that was always encoded by the strain-specific lgs gene. Two of the strains were used to inoculate chickens maintained in a protected environment which were Lactobacillus-free prior to inoculation. DNAs and RNAs extracted from the digesta of the chickens were used for PCR and reverse transcription-PCR, respectively, to demonstrate the presence and transcription of lgs genes in vivo. In both cases, only the strain-specific gene was transcribed. Both of the strains adhered to the crop epithelium, consistent with published data predicting that S-proteins of lactobacilli are adhesins. The results of this study provide a basis for the investigation of gene duplication and sequence variation as mechanisms by which bacterial strains of the same species can share the same habitat.

K.E.H. and L.L.G. contributed equally to this study.

Present address: Institut Rosell Research and Development, 6100 Royalmount Av., Montreal, Quebec H4P 2R2, Canada.

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