Biomaterial Database

Construction of a broad-host-range kanamycin-resistant plasmid vector. 1983

R Roychoudhury, and A Lam

A new 10.2-kb plasmid, pIRL2, was constructed by using a 2140-bp DNA fragment carrying the Kmr gene and BamHI cohesive ends. These BamHI cohesive ends were used to trap the replicating DNA fragment from a partial Sau3A digest of the plasmid R300B. The plasmid contains unique EcoRI, SstI, HindIII, SmaI, SalI, and XhoI sites. These sites can be used as cloning sites without the loss of Kmr. A unique BglII site can be used as a cloning site by insertional inactivation of the Kmr structural gene, coding for neomycin phosphotransferase type II. The new plasmid carries the Sur and Smr genes of R300B. The direction of transcription from the neo promoter is clockwise, the same as that from the sul promoter. The plasmid retains the broad-host-range function of R300B, and thus it may be used for gene cloning in Rhizobium and Agrobacterium for genetic engineering of plant cells.

UI	MeSH Term	Description	Entries
D007612	Kanamycin	Antibiotic complex produced by Streptomyces kanamyceticus from Japanese soil. Comprises 3 components: kanamycin A, the major component, and kanamycins B and C, the minor components.	Kanamycin A,Kanamycin Sulfate,Kantrex
D011815	R Factors	A class of plasmids that transfer antibiotic resistance from one bacterium to another by conjugation.	R Factor,R Plasmid,R Plasmids,Resistance Factor,Resistance Factors,Factor, R,Factor, Resistance,Factors, R,Factors, Resistance,Plasmid, R,Plasmids, R
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
D004261	DNA Replication	The process by which a DNA molecule is duplicated.	Autonomous Replication,Replication, Autonomous,Autonomous Replications,DNA Replications,Replication, DNA,Replications, Autonomous,Replications, DNA
D004262	DNA Restriction Enzymes	Enzymes that are part of the restriction-modification systems. They catalyze the endonucleolytic cleavage of DNA sequences which lack the species-specific methylation pattern in the host cell's DNA. Cleavage yields random or specific double-stranded fragments with terminal 5'-phosphates. The function of restriction enzymes is to destroy any foreign DNA that invades the host cell. Most have been studied in bacterial systems, but a few have been found in eukaryotic organisms. They are also used as tools for the systematic dissection and mapping of chromosomes, in the determination of base sequences of DNAs, and have made it possible to splice and recombine genes from one organism into the genome of another. EC 3.21.1.	Restriction Endonucleases,DNA Restriction Enzyme,Restriction Endonuclease,Endonuclease, Restriction,Endonucleases, Restriction,Enzymes, DNA Restriction,Restriction Enzyme, DNA,Restriction Enzymes, DNA
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
D013045	Species Specificity	The restriction of a characteristic behavior, anatomical structure or physical system, such as immune response; metabolic response, or gene or gene variant to the members of one species. It refers to that property which differentiates one species from another but it is also used for phylogenetic levels higher or lower than the species.	Species Specificities,Specificities, Species,Specificity, Species