Cloning and expression of a mureinolytic enzyme from the mycobacteriophage TM4.
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.
Henry
Marine
M
Department of Biological Sciences, Cork Institute of Technology, Cork, Ireland.
Begley
Máire
M
Neve
Horst
H
Maher
Fiona
F
Ross
Reynolds Paul
RP
McAuliffe
Olivia
O
Coffey
Aidan
A
O'Mahony
Jim M
JM
D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D015870:Gene Expression; D008969:Molecular Sequence Data; D009160:Mycobacteriophages / Q000201:enzymology* / Q000235:genetics / Q000502:physiology; D010457:Peptidoglycan / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D014759:Viral Envelope Proteins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism
D010457:Peptidoglycan; D014759:Viral Envelope Proteins