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Microbiology 155 (2009), 2683-2693; DOI  10.1099/mic.0.027748-0
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Microbiology 155 (2009), 2683-2693; DOI  10.1099/mic.0.027748-0
© 2009 Society for General Microbiology

Comparison of Bacteroides thetaiotaomicron and Escherichia coli 16S rRNA gene expression signals

Matthew D. Mastropaolo{dagger},{ddagger}, Mary L. Thorson{dagger} and Ann M. Stevens

Department of Biological Sciences, Virginia Tech, 219 Life Sciences 1, Washington Street, Blacksburg, VA 24061-0910, USA

There are barriers to cross-expression of genes between Bacteroides spp. and Escherichia coli. In this study, a lux-based reporter system was developed for Bacteroides and used to compare the promoter structure and function of a Bacteroides thetaiotaomicron 4001 (BT4001) 16S rRNA promoter with those of E. coli in vivo. Analysis of the BT4001 sequences upstream of the 16S rRNA gene revealed the same overall structure known for E. coli 16S rRNA promoters in that there were two promoters separated by ~150 bp. However, the BT4001 16S rRNA promoter contains the proposed Bacteroides –7 and –33 consensus sequences instead of the E. coli –10 and –35 consensus sequences. The biological activity of various configurations of the BT4001 16S rRNA promoter was analysed. Experiments pairing the BT4001 16S rRNA promoter with an E. coli RBS, and vice-versa, confirmed that gene expression between the two species is restricted at the level of transcription. In Bacteroides, a difference in translation initiation also appears to limit expression of foreign genes.

Correspondence
Ann M. Stevens
ams{at}vt.edu

Abbreviations: BT4001, Bacteroides thetaiotaomicron 4001; CAT, chloramphenicol acetyltransferase; RLU, relative light units

{dagger}These authors contributed equally to this work.

{ddagger}Present address: Center for Adaptation Genetics and Drug Resistance, Department of Molecular Biology and Microbiology, Tufts University School of Medicine, 136 Harrison Ave, Boston, MA 02111, USA.

A supplementary table, listing primers used in this study, is available with the online version of this paper.






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