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

  • K. Brooks Low
  • Sean R. Murray
  • John Pawelek
  • David BermudesEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1409)


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.

Key words

Salmonella Tumor-targeting Antitumor efficacy msbB Myristoylation Lipid A somA Suwwan pmrAC zwf Suppressor analysis Epistasis Pseudo-reversion Tolerance acquisition Compensatory mutations Rescue mutations Chemical conditionality 



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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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • K. Brooks Low
    • 1
  • Sean R. Murray
    • 2
    • 3
  • John Pawelek
    • 4
  • David Bermudes
    • 2
    • 3
    Email author
  1. 1.Department of Therapeutic Radiology, School of MedicineYale UniversityNew HavenUSA
  2. 2.Biology DepartmentCalifornia State University, NorthridgeNorthridgeUSA
  3. 3.Interdisciplinary Research Institute for the Sciences (IRIS)California State University, NorthridgeNorthridgeUSA
  4. 4.Department of Dermatology, School of MedicineYale UniversityNew HavenUSA

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