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Applied and Environmental Microbiology, February 2008, p. 987-993, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02514-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

Genomic Differences between Fibrobacter succinogenes S85 and Fibrobacter intestinalis DR7, Identified by Suppression Subtractive Hybridization{triangledown} ,{dagger}

M. Qi,1 K. E. Nelson,2 S. C. Daugherty,2 W. C. Nelson,2 I. R. Hance,2 M. Morrison,3 and C. W. Forsberg1*

Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada,1 The J. Craig Venter Institute, Rockville, Maryland,2 Department of Animal Science, The Ohio State University, Columbus, Ohio3

Received 7 November 2007/ Accepted 13 December 2007

Fibrobacter is a highly cellulolytic genus commonly found in the rumen of ruminant animals and cecum of monogastric animals. In this study, suppression subtractive hybridization was used to identify the genes present in Fibrobacter succinogenes S85 but absent from F. intestinalis DR7. A total of 1,082 subtractive clones were picked, plasmids were purified, and inserts were sequenced, and the clones lacking homology to F. intestinalis were confirmed by Southern hybridization. By comparison of the sequences of the clones to one another and to those of the F. succinogenes genome, 802 sequences or 955 putative genes, comprising approximately 409 kb of F. succinogenes genomic DNA, were identified that lack similarity to those of F. intestinalis chromosomal DNA. The functional groups of genes, including those involved in cell envelope structure and function, energy metabolism, and transport and binding, had the largest number of genes specific to F. succinogenes. Low-stringency Southern hybridization showed that at least 37 glycoside hydrolases are shared by both species. A cluster of genes responsible for heme, porphyrin, and cobalamin biosynthesis in F. succinogenes S85 was either missing from or not functional in F. intestinalis DR7, which explains the requirement of vitamin B12 for the growth of the F. intestinalis species. Two gene clusters encoding NADH-ubiquinone oxidoreductase subunits probably shared by Fibrobacter genera appear to have an important role in energy metabolism.

* Corresponding author. Mailing address: Department of Molecular and Cellular Biology, University of Guelph, Guelph, Ontario, Canada N1G 2W1. Phone: (519) 824-4120, ext. 53433. Fax: (519) 837-1802. E-mail: cforsber{at}uoguelph.ca

{triangledown} Published ahead of print on 21 December 2007.

{dagger} Supplemental material for this article may be found at http://aem.asm.org/.

Applied and Environmental Microbiology, February 2008, p. 987-993, Vol. 74, No. 4
0099-2240/08/$08.00+0     doi:10.1128/AEM.02514-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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