RESUMO
A method of efficient purification of the extremely instable virions of the temperate bacteriophage ZF40 of Erwinia carotovora was proposed in the work. The stage-by-stage centrifugation in mentrizamide and caesium chloride gradients permitted to obtain highly purified phage preparations which were used for studying the protein composition of the studied erwiniophage. It was established that the buoyant density of the phage ZF40 was 1.23 g/cm3, 1.49 g/cm3 and 1.27 g/cm3 in the gradients of metrizamide, caesium chloride and caesium sulphate, respectively. By the polypeptide composition of the virion the phage ZF40 has been included in the group of P2-like phages.
Assuntos
Myoviridae/química , Pectobacterium carotovorum/virologia , DNA Viral/análise , Eletroforese em Gel de Poliacrilamida , Lisogenia , Mutação , Myoviridae/genética , Myoviridae/isolamento & purificação , RNA Viral/análiseRESUMO
It was shown that temperate bacteriophage ZF40 of Erwinia carotovora can perform generalized transduction of plasmid pKM 101. The antibiotic-resistance marker transfer is coordinated with the fact of cyclic permutation of the phage genom. The presented results create preconditions for further use of bacteriophage ZF40 as a convenient instrument for genetic study of E. carotovora.
Assuntos
Bacteriófagos/genética , Pectobacterium carotovorum/genética , Plasmídeos/genética , Transdução Genética , DNA Viral/análise , Eletroforese em Gel de Poliacrilamida , Lisogenia , Pectobacterium carotovorum/virologiaRESUMO
It has been shown that the temperate bacteriophage ZF40 Erwinia carotovora combines in it two kinds of intragenome heterogeneity--cyclic permutation and cohesive end structures. The latter reassociate more quickly than analogous cos-sites of the phage lambda Escherichia coli. A series of clear-mutants was found after the temperature inactivation of the phage particles; they are the components of the complementation group being involved in the repressor gene. One of the mutants, ZF40c(5/5), was identified as the virulent one. It has been established that the character of mutation in temperature clear-mutants changes towards its discreteness. Due to the uniqueness of the intragenome organization the molecular-genetic investigations of the processes of replication, site-specific recombination and packing of DNA of the phage ZF40 E. carotovora are of great scientific interest.
