Cloning and expression of a Staphylococcus aureus gene encoding a peptidoglycan hydrolase activity.
A gene of Staphylococcus aureus PS47 encoding lytic activity was cloned and expressed in Escherichia coli. Deletion analysis of a recombinant plasmid carrying a 7.4-kilobase-pair fragment (kbp) of S. aureus DNA suggested that the gene was located within a 2.5-kbp EcoRI-XbaI fragment. Analysis of extracts of E. coli harboring recombinant plasmids on denaturing polyacrylamide gels containing purified cell walls of S. aureus showed a clearing zone by a polypeptide of apparent Mr 23,000. The release of dinitrophenylalanine but not reducing groups from purified cell walls by a cell extract of recombinant E. coli suggested that we had cloned an N-acetylmuramyl-L-alanine amidase.
Jayaswal
R K
RK
Department of Biological Sciences, Illinois State University, Normal 61761.
Lee
Y I
YI
Wilkinson
B J
BJ
D002473:Cell Wall / Q000648:ultrastructure; D003001:Cloning, Molecular / Q000379:methods; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D007700:Kinetics; D009238:N-Acetylmuramoyl-L-alanine Amidase / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism; D015183:Restriction Mapping; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics*
D004269:DNA, Bacterial; D007527:Isoenzymes; D009238:N-Acetylmuramoyl-L-alanine Amidase