Expression of the S. aureus hysA gene in S. carnosus from a modified E. coli-staphylococcal shuttle vector.
We have modified an E. coli-staphylococcal shuttle vector for use in the general cloning and expression of genes from pathogenic staphylococci in Staphylococcus carnosus. As S. carnosus is non-pathogenic, this expression system will facilitate the study of the roles of individual gene products in the disease process. To evaluate the use of this expression system, a DNA fragment containing the Staphylococcus aureus hyaluronate lyase (hysA) gene was cloned into the modified vector, pNW21, and introduced into S. carnosus. Hyaluronate lyase was both produced and secreted by S. carnosus. In addition, the secreted HysA protein was enzymatically active, as determined using a zymographic assay.
Williams
R J
RJ
Cellular Microbiology Research Group, Division of Surgical Sciences, Eastman Dental Institute, UK.
Nair
S P
SP
Henderson
B
B
Holland
K T
KT
Ward
J M
JM
D001483:Base Sequence; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D015870:Gene Expression; D005818:Genetic Engineering; D005822:Genetic Vectors / Q000235:genetics*; D011133:Polysaccharide-Lyases / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D013210:Staphylococcus / Q000235:genetics*; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics* / Q000472:pathogenicity; D014158:Transcription, Genetic; D014774:Virulence / Q000235:genetics
D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase