Biomaterial Database

Isolation and characterization of deletion derivatives of pDL282, an Actinobacillus actinomycetemcomitans/Escherichia coli shuttle plasmid. 1994

P K Sreenivasan, and P Fives-Taylor

Markey Center for Molecular Genetics, College of Medicine, University of Vermont, Burlington 05405.

Recent reports have described the construction of several shuttle plasmids (D.J. LeBlanc, L. L. Lee, A. Al-Jaibat, P. K. Sreenivasan, and P. M. Fives-Taylor, Oral Micro. Immunol. (1993) 8, 94-99) and the development of an efficient transformation system for Actinobacillus actinomycetemcomitans (P. K. Sreenivasan, D. J. LeBlanc, L. L. Lee, and P. M. Fives-Taylor, Infect. Immun. (1991) 59, 4621-4627), a gram-negative bacterium implicated in human periodontal disease. This report presents results from further studies on pDL282, an A. actinomycetemcomitans-Escherichia coli shuttle plasmid. A. actinomycetemcomitans containing pDL282 lost the plasmid at a rapid rate when cultured in antibiotic free medium. Intact pDL282 was maintained for 20 generations or more when the host cells were grown in the presence of ampicillin, or ampicillin plus spectinomycin. However, prolonged incubation in the presence of spectinomycin only resulted in the emergence of one or the other of two unique deletion derivatives of pDL282, designated pPK1 and pPK2. Whereas A. actinomycetemcomitans was efficiently transformed with pDL282 (5.7 kb), pPK1 (3.6 kb), and pPK2 (2.5 kb), E. coli was transformed only by the two largest species. Like the parent molecule, pPK1 and pPK2 were rapidly lost from A. actinomycetemcomitans hosts in the absence of antibiotic selection. Neither pPK1 nor pPK2 suffered any further deletions following prolonged cultivation in the presence of spectinomycin. The minimal replicon of pVT736-1, the A. actinomycetemcomitans-derived plasmid component of pDL282, was located within a 1400 bp DNA fragment of pPK1 and pPK2.(ABSTRACT TRUNCATED AT 250 WORDS)

UI	MeSH Term	Description	Entries
D010957	Plasmids	Extrachromosomal, usually CIRCULAR DNA molecules that are self-replicating and transferable from one organism to another. They are found in a variety of bacterial, archaeal, fungal, algal, and plant species. They are used in GENETIC ENGINEERING as CLONING VECTORS.	Episomes,Episome,Plasmid
D002876	Chromosomes, Bacterial	Structures within the nucleus of bacterial cells consisting of or containing DNA, which carry genetic information essential to the cell.	Bacterial Chromosome,Bacterial Chromosomes,Chromosome, Bacterial
D004269	DNA, Bacterial	Deoxyribonucleic acid that makes up the genetic material of bacteria.	Bacterial DNA
D004591	Electrophoresis, Polyacrylamide Gel	Electrophoresis in which a polyacrylamide gel is used as the diffusion medium.	Polyacrylamide Gel Electrophoresis,SDS-PAGE,Sodium Dodecyl Sulfate-PAGE,Gel Electrophoresis, Polyacrylamide,SDS PAGE,Sodium Dodecyl Sulfate PAGE,Sodium Dodecyl Sulfate-PAGEs
D004926	Escherichia coli	A species of gram-negative, facultatively anaerobic, rod-shaped bacteria (GRAM-NEGATIVE FACULTATIVELY ANAEROBIC RODS) commonly found in the lower part of the intestine of warm-blooded animals. It is usually nonpathogenic, but some strains are known to produce DIARRHEA and pyogenic infections. Pathogenic strains (virotypes) are classified by their specific pathogenic mechanisms such as toxins (ENTEROTOXIGENIC ESCHERICHIA COLI), etc.	Alkalescens-Dispar Group,Bacillus coli,Bacterium coli,Bacterium coli commune,Diffusely Adherent Escherichia coli,E coli,EAggEC,Enteroaggregative Escherichia coli,Enterococcus coli,Diffusely Adherent E. coli,Enteroaggregative E. coli,Enteroinvasive E. coli,Enteroinvasive Escherichia coli
D006801	Humans	Members of the species Homo sapiens.	Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000188	Actinobacillus	A genus of PASTEURELLACEAE described as gram-negative, nonsporeforming, nonmotile, facultative anaerobes. Most members are found both as pathogens and commensal organisms in the respiratory, alimentary, and genital tracts of animals.
D000668	Ampicillin Resistance	Nonsusceptibility of a microbe to the action of ampicillin, a penicillin derivative that interferes with cell wall synthesis.	Ampicillin Resistances,Resistance, Ampicillin,Resistances, Ampicillin
D014170	Transformation, Genetic	Change brought about to an organisms genetic composition by unidirectional transfer (TRANSFECTION; TRANSDUCTION, GENETIC; CONJUGATION, GENETIC, etc.) and incorporation of foreign DNA into prokaryotic or eukaryotic cells by recombination of part or all of that DNA into the cell's genome.	Genetic Transformation,Genetic Transformations,Transformations, Genetic
D015183	Restriction Mapping	Use of restriction endonucleases to analyze and generate a physical map of genomes, genes, or other segments of DNA.	Endonuclease Mapping, Restriction,Enzyme Mapping, Restriction,Site Mapping, Restriction,Analysis, Restriction Enzyme,Enzyme Analysis, Restriction,Restriction Enzyme Analysis,Analyses, Restriction Enzyme,Endonuclease Mappings, Restriction,Enzyme Analyses, Restriction,Enzyme Mappings, Restriction,Mapping, Restriction,Mapping, Restriction Endonuclease,Mapping, Restriction Enzyme,Mapping, Restriction Site,Mappings, Restriction,Mappings, Restriction Endonuclease,Mappings, Restriction Enzyme,Mappings, Restriction Site,Restriction Endonuclease Mapping,Restriction Endonuclease Mappings,Restriction Enzyme Analyses,Restriction Enzyme Mapping,Restriction Enzyme Mappings,Restriction Mappings,Restriction Site Mapping,Restriction Site Mappings,Site Mappings, Restriction