New suicide vector for gene replacement in yersiniae and other gram-negative bacteria. 1993

E Skrzypek, and P L Haddix, and G V Plano, and S C Straley
Department of Microbiology and Immunology, Chandler Medical Center, University of Kentucky, Lexington 40536-0084.

A suicide vector named pUK4134 was constructed to enlarge the repertoire of vectors available for allelic exchange of mutated sequences in gram-negative bacteria. This plasmid combines the properties of two previously described plasmid vectors, pJM703.1 and pRTP1. pUK4134 is a suicide vector, carrying the origin of replication of the plasmid R6K and thus requiring the product of the pir gene for its stable maintenance. The rpsL gene encoding Escherichia coli ribosomal protein S12 confers streptomycin sensitivity on streptomycin-resistant strains and provides a positive selection for bacteria that have lost the plasmid following allelic exchange. The bla gene provides for selection by Apr. Other features are a unique EcoRV cloning site, oriT of plasmid RK2, and the bacteriophage lambda cos sequence. This vector was successfully used several times to carry out allelic exchange in Yersinia pestis.

UI MeSH Term Description Entries
D010582 Bacteriophage lambda A temperate inducible phage and type species of the genus lambda-like viruses, in the family SIPHOVIRIDAE. Its natural host is E. coli K12. Its VIRION contains linear double-stranded DNA with single-stranded 12-base 5' sticky ends. The DNA circularizes on infection. Coliphage lambda,Enterobacteria phage lambda,Phage lambda,lambda Phage
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
D012093 Replicon Any DNA sequence capable of independent replication or a molecule that possesses a REPLICATION ORIGIN and which is therefore potentially capable of being replicated in a suitable cell. (Singleton & Sainsbury, Dictionary of Microbiology and Molecular Biology, 2d ed) Replication Unit,Replication Units,Replicons,Unit, Replication,Units, Replication
D003001 Cloning, Molecular The insertion of recombinant DNA molecules from prokaryotic and/or eukaryotic sources into a replicating vehicle, such as a plasmid or virus vector, and the introduction of the resultant hybrid molecules into recipient cells without altering the viability of those cells. Molecular Cloning
D005798 Genes, Bacterial The functional hereditary units of BACTERIA. Bacterial Gene,Bacterial Genes,Gene, Bacterial
D005822 Genetic Vectors DNA molecules capable of autonomous replication within a host cell and into which other DNA sequences can be inserted and thus amplified. Many are derived from PLASMIDS; BACTERIOPHAGES; or VIRUSES. They are used for transporting foreign genes into recipient cells. Genetic vectors possess a functional replicator site and contain GENETIC MARKERS to facilitate their selective recognition. Cloning Vectors,Shuttle Vectors,Vectors, Genetic,Cloning Vector,Genetic Vector,Shuttle Vector,Vector, Cloning,Vector, Genetic,Vector, Shuttle,Vectors, Cloning,Vectors, Shuttle
D006090 Gram-Negative Bacteria Bacteria which lose crystal violet stain but are stained pink when treated by Gram's method. Gram Negative Bacteria
D000091243 Ribosomal Protein S9 It is a protein that encodes a RIBOSOMAL PROTEIN which is a component of the 40S subunit. It belongs to the S4P family of ribosomal proteins. S9 Ribosomal Protein,Protein S9, Ribosomal,Protein, S9 Ribosomal,S9, Ribosomal Protein
D015007 Yersinia A genus of gram-negative, facultatively anaerobic rod- to coccobacillus-shaped bacteria that occurs in a broad spectrum of habitats.
D029968 Escherichia coli Proteins Proteins obtained from ESCHERICHIA COLI. E coli Proteins

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