Abstract
Bacteriolytic anti-cancer therapies employ attenuated bacterial strains that selectively proliferate within tumors. Clostridium novyi-NT spores represent one of the most promising of these agents, as they generate potent anti-tumor effects in experimental animals. We have determined the 2.55-Mb genomic sequence of C. novyi-NT, identifying a new type of transposition and 139 genes that do not have homologs in other bacteria. The genomic sequence was used to facilitate the detection of transcripts expressed at various stages of the life cycle of this bacterium in vitro as well as in infections of tumors in vivo. Through this analysis, we found that C. novyi-NT spores contained mRNA and that the spore transcripts were distinct from those in vegetative forms of the bacterium.
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Acknowledgements
The authors thank Peter Setlow for his insightful suggestions about the nature of spore mRNA, Rachel Green for her comments on rRNA fragmentation, Anca Segall for her comments on transposition, and Tanja Davidsen, Michelle Gwinn Giglio and Nikhat Zafar of TIGR for their expert assistance with the genomic bioinformatic analysis. The authors also thank Brent Ewing, Phil Green and David Gordon for kindly providing the Phred/Phrap and Consed software package. The B. subtilis strains were generously provided by Peter Mullany. This work was supported by the Virginia and D.K. Ludwig Fund for Cancer Research, the Commonwealth Foundation, the Miracle Foundation, National Science Foundation grant DMS034211 and National Institutes of Health grant CA062924.
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Supplementary information
Supplementary Fig. 1
rRNA species. (PDF 110 kb)
Supplementary Fig. 2
RT-PCR confirmation of selected microarray data. (PDF 143 kb)
Supplementary Fig. 3
Transcript abundance in denuded spores. (PDF 134 kb)
Supplementary Table 1
Integrated database of C. novyi-NT genome and transcriptomes. (XLS 1009 kb)
Supplementary Table 2
Coding sequences related to mobile elements. (XLS 38 kb)
Supplementary Table 3
Hypothetical CDS unique to C. novyi-NT. (PDF 54 kb)
Supplementary Table 4
Functional classification of C. novyi-NT genes. (PDF 75 kb)
Supplementary Table 5
Other bacteria harboring homologs of the C. novyi-NT transposase. (PDF 82 kb)
Supplementary Table 6
Functional classification of C. novyi-NT genes expressed in specific growth phases. (PDF 68 kb)
Supplementary Table 7
Genes expressed preferentially in different growth phases. (XLS 104 kb)
Supplementary Table 8
Fifthy genes with transcripts most abundant in spores. (XLS 328 kb)
Supplementary Table 9
Genes transcribed during the final stages of sporulation. (XLS 214 kb)
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Bettegowda, C., Huang, X., Lin, J. et al. The genome and transcriptomes of the anti-tumor agent Clostridium novyi-NT. Nat Biotechnol 24, 1573–1580 (2006). https://doi.org/10.1038/nbt1256
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DOI: https://doi.org/10.1038/nbt1256
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