RRC ID 29771
著者 Lovett MA, Helinski DR.
タイトル Method for the isolation of the replication region of a bacterial replicon: construction of a mini-F'kn plasmid.
ジャーナル J Bacteriol
Abstract A purified fragment of deoxyribonucleic acid (DNA) that determines resistance to kanamycin and is incapable of self-replication was used to select a self-replicating fragment from an EcoRI endonuclease digest of the sex factor F'lac. This F'lac fragment, exhibiting a molecular weight of 6 X 10(6), carries the genes essential for maintenance of the F replicon in Escherichia coli cells. The constructed mini-F'km plasmid also retains the incompatibility properties of the parent F'lac plasmid. Large amounts of the kanamycin resistance fragment of a molecular weight of 4.5 X 10(6) with an EcoRI-cleaved, self-replicating derivative of colicinogenic plasmid E1 that has a molecular weight of 2.2 X 10(6), The recombinant plasmid is able to replicate extensively in E. coli in medium containing chloramphenicol, and, therefore, large quantities of this plasmid DNA can be obtained. The substantial difference in size between the two fragments in the recombinant plasmid greatly facilitates their separation by preparative agarose gel electrophoresis.
巻・号 127(2)
ページ 982-7
公開日 1976-8-1
DOI 10.1128/jb.127.2.982-987.1976
PMID 783124
PMC PMC233008
MeSH Chloramphenicol / pharmacology Colicins / pharmacology Conjugation, Genetic* DNA Replication DNA Restriction Enzymes / metabolism DNA, Bacterial / biosynthesis DNA, Bacterial / isolation & purification* Drug Resistance, Microbial Escherichia coli / analysis* Escherichia coli / drug effects Escherichia coli / metabolism F Factor* Kanamycin / pharmacology Molecular Weight
IF 3.006
リソース情報
原核生物(大腸菌) ME6232