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Content archived on 2024-04-15

MOLECULAR BIOLOGY OF BACILLUS SUBTILIS BACTERIOPHAGES

Objective


The sizing gene (gene 6) of Bacillus subtilis (Gram positive) bacteriophage SPP1 and several mutant alleles were characterized with respect to cloning frequency. For the first time a cistron which affects the postulated headful sizing function was characterized. An in vivo assay to test the undersizing activity of gene 6 mutant alleles was also developed.

Analysis of the gene 6 gene product (Gp6) placed it in a group of polypeptides previously described in the literature for other phages as the portal vertex proteins. No primary structure similarity between any polypeptide of this group could be detected.
The preliminary genetic and functional analysis of cistron 6 together with the biochemical and structural data concerning its gene product which were obtained during the present study provide powerful tools for a systematic study of the portal protein function in deoxyribonucleic acid (DNA) sizing during packaging.

The lysogeny in group III B. subtilis bacteriophage was studied. Analysis of the superinfection immunity specificity (ie the refractory state of lysogenic bacteria to infection with phage of the carried type) showed that 2 main homoimmune subgroups can be distinguished: subgroup I, including SPbeta, SPR, Z, IG1, IG3, IG4, H2 and E; and subgroup II, formed by phi3T and p11. This division possibly reflects differences in the way the phages from each subgroup maintain their lysogenic state.
The research provided a probe to map the SPbeta genomic region responsible for superinfection immunity, creating, for the first time, a feasible experimental way to access this problem in B. subtilis group III temperate bacteriophages. Furthermore, it shows a particularly interesting relationship between the bacterial chromosome structure and the genome of phages that are able to invade it.

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FUNDACAO CALOUSTE GULBENKIAN
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