RESUMO
Bacteriophage BFK20 was isolated from a Brevibacterium flavum strain that had become contaminated during industrial fermentation. BFK20 has a polyhedral head 50 nm wide and a non-contractile tail 200 nm long and 10 nm in diameter. The genome of this bacteriophage consists of a linear double stranded DNA molecule of 44-45 kb with cohesive ends. The capsid of phage BFK20 contains nine polypeptides with molecular masses from 22.0-108.0 kDa. BFK20 DNA was used as a donor for fragments carrying promoters and transcription-terminators.
Assuntos
Bacteriófagos/genética , Brevibacterium , Bacteriófagos/metabolismo , Bacteriófagos/ultraestrutura , Eletroforese em Gel de Poliacrilamida , Genoma Viral , Microscopia Eletrônica , Regiões Promotoras Genéticas , Mapeamento por Restrição , Transcrição GênicaRESUMO
We constructed a promoter-probe vector, pJUP05, for brevibacteria and Escherichia coli based on the promoterless neomycin-resistance (neoR) gene from Tn5. This gene confers resistance to the aminoglycosides, kanamycin and neomycin. The promoter of the neoR gene was deleted and replaced by a suitable multiple cloning site. There are translation stop codons in all three reading frames upstream from the neoR gene. The plasmid contains functional origins of DNA replication for both brevibacteria and E. coli, and permits selection for chloramphenicol- and/or ampicillin-resistance markers.
