Restriction map of bacteriophage SP02: ordering the SacI fragments and identification of the DNA termini |
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| Authors: | Y Yoneda S Graham T Evans F E Young |
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| Affiliation: | 1. Department of Microbiology and Immunology, Virginia Commonwealth University, Richmond, VA 23298, USA;2. Departments of Oral Biology, and Microbiology and Immunology, Dental Research Institute, University of Michigan, Schools of Medicine and Dentistry, Ann Arbor, MI 48109 U.S.A. |
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| Abstract: | A plasmid that is able to replicate in both Escherichia coli and Streptococcus sanguis has been constructed by the in vitro joining of the pACYC184 (Cmr Tcr) and pVA749 (Emr) replicons. This plasmid, designated pVA838, is 9.2 kb in size and expresses Emr in both E. coli and S. sanguis. Its Cmr marker is expressed only in E. coli and may be inactivated by addition of DNA inserts at its internal EcoRI or PvuII sites. The pVA838 molecule also contains unique SalI, SphI, BamHI, NruI and XbaI cleavage sites suitable for molecular cloning. pVA838 may be amplified in E. coli but not in S. sanguis. We have used the pVA838 plasmid as a shuttle vector to clone streptococcai plasmid fragments in E. coli. Such chimeras isolated from E. coli were readily introduced into S. sanguis by transformation. |
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| Keywords: | Recombinant DNA streptococcal shuttle plasmid pACYC184 pVA749 pVA838 CCC covalently closed circular chloramphenicol resistance chloramphenicol sensitivity erythromycin resistance erythromycin sensitivity kb kilobase pairs SDS sodium dodecyl sulfate tetracycline resistance tetracycline sensitivity :: novel joint [] indicates plasmid-carrier state |
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