Cloning and expression of a mureinolytic enzyme from the mycobacteriophage TM4
Document Type
Article
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
Disciplines
Biology
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
Publication Details
FEMS Microbiology Letters