Abstract
The plasmid pT181 of Staphylococcus aureus consists of 4437 base pairs and encodes resistance to tetracycline. Initiation of pT181 replication specifically requires the plasmid-encoded repC protein. An in vitro system has been shown to carry out semiconservative replication of pT181 and its derivative plasmids (Khan, S A., Carleton, S. M., and Novick, R. P. (1981) Proc. Natl. Acad. Sci. U. S. A. 78, 4902-4906). We have used this replication assay to isolate repC protein, which was purified to near homogeneity. The repC gene was cloned into the pKJB825 plasmid that contains the phage lambda temperature-sensitive repressor gene, cI857, and the rightward promoter, PR. Upon temperature induction, Escherichia coli clones containing the recombinant plasmid overproduced repC protein, which was purified in significant quantities. The molecular weight of repC protein under denaturing conditions is 38,000, which is consistent with the size predicted from the DNA sequence data. Presence of repC protein was absolutely essential for the initiation of replication of pT181 and its derivatives in vitro.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
