Cloning and expression of a mureinolytic enzyme from the mycobacteriophage TM4

Authors

Marine Henry, Cork Institute of Technology
Máire Begley, University College Cork
Horst Neve, Max Rubner-Institut, Germany
Fiona Maher, Department of Biological Sciences, Cork Institute of Technology, Cork, IrelandFollow
Reynolds Paul Ross, Moorepark Food Research Centre
Olivia McAuliffe, Moorepark Food Research Centre
Aidan Coffey, Department of Biological Sciences, Cork Institute of Technology, Cork, IrelandFollow
Jim M. O'Mahony, Department of Biological Sciences, Cork Institute of Technology, Cork, IrelandFollow

Document Type

Article

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

Disciplines

Biology

Publication Details

FEMS Microbiology Letters

Abstract

In this study, we describe the characterization, cloning, expression and purification of the lysin A gene of the mycobacteriophage TM4. The gene TM4-gp29 (gp29) is a 1644-bp gene that codes for a 58.6-kDa protein and contains peptidoglycan recognition protein, Zn-binding and amidase catalytic domains. The gene was cloned into Escherichia coli using the 'His-Tag' pQE60 vector. After affinity chromatography-mediated purification, the protein was concentrated and visualized using sodium dodecyl sulphate polyacrylamide gel electrophoresis. Evidence of peptidoglycan-degrading activity was observed initially by a chloroform assay and later by conventional zymogram analysis. © 2010 Federation of European Microbiological Societies.

Recommended Citation

Henry, Marine; Begley, Máire; Neve, Horst; Maher, Fiona; Ross, Reynolds Paul; McAuliffe, Olivia; Coffey, Aidan; and O'Mahony, Jim M., "Cloning and expression of a mureinolytic enzyme from the mycobacteriophage TM4" (2010). Department of Biological Sciences Publications [online].
Available at: https://doi.org/10.1111/j.1574-6968.2010.02080.x