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Studying Gene Induction of Glycopeptide Resistance Using Gene Swapping

  • Hee-Jeon Hong
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 642)

Abstract

Gene swapping is a simple but effective genetic tool for characterizing the functioning of a gene, where the gene in question is known to fulfil a distinctive biological role in the cell. VanS is a sensor kinase which, in conjunction with its cognate response regulator VanR, triggers resistance to vancomycin. One of the most important questions yet to be answered in the study of vancomycin resistance is the nature of the specific ligand recognized by the VanS sensor. A “VanRS-swap” experiment between two glycopeptide-resistant Streptomyces species known to exhibit differing responses to inducer molecules can investigate whether inducer specificity is determined solely by differences between the amino acid sequences of the VanRS two-component systems present, or by inherent differences in cell wall structure and biosynthesis between the strains. Results from such experiments demonstrate that inducer specificity is determined by the origin of the VanRS proteins and provides useful circumstantial evidence that the VanS effector ligand is the drug itself, and not an intermediate in cell wall biosynthesis that may accumulate as a result of drug action.

Key words

Vancomycin resistance Glycopeptides S. coelicolor S. toyocaensis VanS/VanR signal transduction system Gene swap VanS effector ligand 

Notes

Acknowledgment

The author would like to thank Andrew Hesketh for comments on the manuscript.

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Hee-Jeon Hong
    • 1
  1. 1.Department of BiochemistryUniversity of CambridgeCambridgeUK

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