The SAS proposals for participation in FP6 projects
are listed in blocks according to the FP6 priority theme structure

FP6 priority
1.1.1   Genomics and Biotechnology for Health
Title of the proposal

Bacteriophage BFK20 genome as a model for functional and structural analysis of unknown genes

Slovak Academy of Sciences, Institute of Molecular Biology
Dubravska cesta 21, 842 51 Bratislava, Slovak Republic
Gabriela BUKOVSKA, RNDr. PhD.
+421 2 59307429

Research subject for a potential FP6 project

Research subject for a potential FP6 project: We want to focus on the functional analysis of genes and their coding sequences (open reading frames [ORFs], purification of encoded protein, examination of phenotypes, determination of intracellular localisation and study of interaction with other proteins.We determined and completed whole nucleotide sequence (42 975bp) of our model bacteriophage BFK20, lytic phage of Brevibacterium flavum. We localized 56 open reading frames (ORF) on the corynephage BFK20 genome. Comparing with gene databases reveals a high homology with 10 up to now characterised proteins. Most of others ORFs has unknown function without a significant homology. We want to clarify their biological and biochemical function. Identification and characterisation of the genes, which are essential for phage amplification and studying the mutants containing an essential gene disruption – this will reveal roles of the genes in phage life cycle. By investigation of the replication process it will be postulated a hypothesis of phage DNA replication mechanism. Using our results we want to focus on investigation of gene function, regulation of transcription and protein expression in corynebacteria species.Overproduction of proteins requires a detailed knowledge of the structure and function of transcriptional and translational signals. The stringent mutational and biochemical analysis of selected promoters will be necessary to gain detailed information on the structure-function relationship of promoters (or promoter classes) in corynebacteria and thus a better understanding of gene expression in this organism. Now, the characterisation of the promoter region and corresponding proteins on phage DNA will be a great contribution not only to the study of gene expression, but also an information about corynephage codone usage, transcriptional signals, regulation of transcription in this genera. Identification and characterisation of the genes, which are essential for phage amplification and their disruption will reveal not only their roles in phage life cycle as well as mutual interactions, but also link of the proteins in regulation and transcriptional dependency. From this point of view the study of phage BFK20 and its genome represents an excellent model for widening the knowledge about corynebacteria species.

Recent international cooperation of the research team

Institute of Molecular genetics Academy of Sciences, Praha, Czech republic, Institute of Inherited Metabolic disorders LFUK, Praha, Czech republic.

Proposerīs relevant publications related to the research subject

Ugorcakova J., Bukovska G.,Timko, J.: Construction of the promoter-probe shuttle vectors for Escherichia coli and corynebacteria on the basis of promoterless alfa-amylase gene. Folia Microbiologica 2000, Vol 45, p 114-120,
Raslik I., Bukovska G., Klucar L., Godany A., Timko J.: Cloning and sequence analysis of the ~17 kbp genome fragment from Brevibacterium flavum CCM 251 Bacteriophage BFK20.: Chem. Listy, 2000, vol 94, p. 526
Halgasova N, Bukovska G, Timko J, Kormanec J. Cloning and transcriptional characterization of two sigma factor genes, sigA and sigB, from Brevibacterium flavum. CURR MICROBIOL 43 (4): 249-254, 2001