Home HOME Acta Virologica 2014 Acta Virologica Vol.58, No.2 p.152-159, 2014

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Quarterly,
Founded: 1957
ISSN 0001-723X
E-ISSN 1336-2305

Published in English

Impact Factor = 1.82

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Acta Virologica Vol.58, No.2 p.152-159, 2014

Title: Immunodetection and N-terminal sequencing of DNA replication proteins of bacteriophage BFK20 – lytic phage of Brevibacterium flavum
Author: G. BUKOVSKÁ, N. HALGAŠOVÁ, L. HROMADOVÁ, H. KOŠČOVÁ, M. BUKOVSKÝ

Abstract: Phages are excellent models for studying the mechanism of DNA replication in prokaryotes. Identification of phage proteins involved in phage DNA replication is the first prerequisite for elucidation of the phage replication module. We focused on replication proteins gp41 (a putative helicase from SF2 superfamily), gp43 (a RepA-like protein), and gp44 (a putative DNA polymerase A) of phage BFK20 grown in Brevibacterium flavum. To identify them in the phage-host system, we prepared antibodies to these proteins which were cloned and expressed in Escherichia coli as his-tagged recombinant proteins. After purification to homogeneity the recombinant proteins served for raising specific polyclonal antibodies in mice. Using these antibodies in Western blot analysis the phage proteins gp41, gp43 and gp44 were detected during the phage growth cycle. The proteins gp41 and gp43, prepared from cell lysate by ammonium sulphate precipitation, were N-terminally sequenced and found to contain the sequences N-SVKPRELR-C and N-MLGSTML-C, respectively. This means that gp41 starts with serine but not with common methionine. We consider these findings an initial but important step towards more thorough characterization of replication proteins of phage BFK20.

Keywords: phage BFK20; Brevibacterium flavum; DNA replication; proteins; immunodetection; N-terminal sequence
Year: 2014, Volume: 58, Issue: 2 Page From: 152, Page To: 159
doi:10.4149/av_2014_02_152


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