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Isolation and Analysis of Suppressor Mutations in Tumor-Targeted msbB Salmonella

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Bacterial Therapy of Cancer

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1409))

Abstract

Tumor-targeted Salmonella offers a promising approach to the delivery of therapeutics for the treatment of cancer. The Salmonella strains used, however, must be stably attenuated in order to provide sufficient safety for administration. Approaches to the generation of attenuated Salmonella strains have included deletion of the msbB gene that is responsible for addition of the terminal myristol group to lipid A. In the absence of myristoylation, lipid A is no longer capable of inducing septic shock, resulting in a significant enhancement in safety. However, msbB Salmonella strains also exhibit an unusual set of additional physiological characteristics, including sensitivities to NaCl, EGTA, deoxycholate, polymyxin, and CO2. Suppressor mutations that compensate for these sensitivities include somA, Suwwan, pmrA C, and zwf. We describe here methods for isolation of strains with compensatory mutations that suppress these types of sensitivities and techniques for determining their underlying genetic changes and analysis of their effects in murine tumor models.

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Acknowledgments

This work was supported by start-up funds from the California State University, Northridge College of Mathematics and Science (for DB). KBL, JP, and DB express their admiration for the late Helen Coley Nauts (1907–2001) and appreciation for her meeting with them in April 2000 to discuss the work of her late father William B. Coley. We also thank the anonymous reviewers for their helpful comments.

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Authors and Affiliations

  1. Department of Therapeutic Radiology, School of Medicine, Yale University, New Haven, CT, 06511, USA

    K. Brooks Low

  2. Biology Department, California State University, Northridge, Northridge, CA, 91330-8303, USA

    Sean R. Murray & David Bermudes

  3. Interdisciplinary Research Institute for the Sciences (IRIS), California State University, Northridge, Northridge, CA, 91330-8303, USA

    Sean R. Murray & David Bermudes

  4. Department of Dermatology, School of Medicine, Yale University, New Haven, CT, 06511, USA

    John Pawelek

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  1. K. Brooks Low
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  2. Sean R. Murray
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  3. John Pawelek
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  4. David Bermudes
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Correspondence to David Bermudes .

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Editors and Affiliations

  1. Department of Surgery, AntiCancer Inc, Univ of California – San Diego, San Diego, California, USA

    Robert M Hoffman

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Low, K.B., Murray, S.R., Pawelek, J., Bermudes, D. (2016). Isolation and Analysis of Suppressor Mutations in Tumor-Targeted msbB Salmonella . In: Hoffman, R. (eds) Bacterial Therapy of Cancer. Methods in Molecular Biology, vol 1409. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3515-4_10

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  • DOI: https://doi.org/10.1007/978-1-4939-3515-4_10

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