Using the pRM30 plasmid, an Aps deletion derivative of broad host range plasmid RP4 with integrated new miniMu 5 (11 kb), we followed the transfer of Escherichia coli chromosomal genes to the recipient strain. The miniMu 5-mediated transposition of chromosomal genes occurs onto the plasmid with integrated miniMu 5 rather than onto the "recipient" plasmid pNH602. The plasmid DNA in recipient cells was detected by electrophoresis. One of the acquired hybrid plasmids pTB2 was analyzed genetically and by restriction endodeoxyribonuclease digestion. A structure consisting of miniMu-chromosomal segment-miniMu as a product of Mu-mediated transposition was detected.

• MeSH
• bakteriální chromozomy fyziologie   MeSH
• bakteriální geny * MeSH
• chromozomální delece MeSH
• Escherichia coli genetika   MeSH
• genotyp MeSH
• konjugace genetická MeSH
• křížení genetické MeSH
• plazmidy * MeSH
• transpozibilní elementy DNA * MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• transpozibilní elementy DNA * MeSH

Vector properties of plasmid pNH602, a higher-copy-number deletion mutant of plasmid R6K, were tested by cloning the 6.5 Mg/mol BamHI pSa fragment carrying determinants of resistance to four antibiotics in the unique BamHI site of pNH602. The resulting in vitro constructed recombinant plasmid pNH606 was found to be stable, conjugative, multicopy (20 copies of pNH606 per E. coli chromosome were estimated) and to ensure the increased expression of different genes responsible for the antibiotic resistance. The pSa fragment inserted in the BamHI site of plasmid pNH602 (located in Tn2660) was proved to be transposable to other replicons. Recombinant plasmid pNH606 was analyzed using restriction enzymes BamHI and EcoRI and its physical and genetic map was constructed.

• MeSH
• antibiotická rezistence MeSH
• deoxyribonukleasa BamHI MeSH
• deoxyribonukleasa EcoRI MeSH
• Escherichia coli genetika   MeSH
• genetické vektory * MeSH
• klonování DNA metody   MeSH
• plazmidy * MeSH
• rekombinantní DNA * MeSH
• restrikční enzymy MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• deoxyribonukleasa BamHI MeSH
• deoxyribonukleasa EcoRI MeSH
• rekombinantní DNA * MeSH
• restrikční enzymy MeSH

Formation of a recombinant plasmid designated pNH603 was observed when two plasmids from incompatibility group X, the multicopy plasmid pNH602 (a higher-copy-number deletion derivative of R6K) and the oligocopy plasmid R485, coexisted in a single Escherichia coli cell. According to its size and its restriction endonuclease cleavage pattern, plasmid pNH603 is a true cointegrate of pNH602 and R485. An insertion-sequence-like element coming from plasmid R485 is supposed to mediate the fusion of both replicons. The pNH603 copy number (1-2 per chromosome) indicates that the mechanism of replication of the low-copy-number plasmid is dominant in this cointegrate. No dissociation of pNH603 to parental plasmids was observed even in E. coli K-12 recA+ cells. On the other hand, deletion derivatives of four size classes originate from pNH603 in both recA+ and recA hosts. A miniplasmid designated pNH604, a representative of the most frequent 7 Mg/mol size class, was found, in a low number of copies per host chromosome.

• MeSH
• deoxyribonukleasa BamHI MeSH
• deoxyribonukleasa EcoRI MeSH
• DNA bakterií analýza   izolace a purifikace   ultrastruktura   MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• elektronová mikroskopie MeSH
• Escherichia coli genetika   MeSH
• plazmidy * MeSH
• rekombinace genetická MeSH
• replikace DNA MeSH
• restrikční enzymy MeSH
• transpozibilní elementy DNA MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• deoxyribonukleasa BamHI MeSH
• deoxyribonukleasa EcoRI MeSH
• DNA bakterií MeSH
• restrikční enzymy MeSH
• transpozibilní elementy DNA MeSH