Summary: The temperate bacteriophage VWB integrates into the chromosome of ETH14630 via site-specific integration. Following recombination of the VWB region with the chromosomal sequence, the host-phage junctions and are formed. Nucleotide sequence analysis of and revealed a 45 bp common core sequence. In this 45 bp sequence consists of the 3′ end of a putative tRNA(AGG) gene with a 3′-terminal CCA sequence which is typical for prokaryotic tRNAs. Phage DNA integration restores the putative tRNA(AGG) gene in . However, following recombination the CCA sequence is missing as is the case for most tRNA genes described so far. Adjacent to VWB , an ORF encoding a 427 aa protein was detected. The C-terminal region of this protein shows high similarity to the conserved C-terminal domain of site-specific recombinases belonging to the integrase family. To prove the functionality of this putative integrase gene , an integrative vector pKT02 was constructed. This vector consists of a 2·3 kb III-l restriction fragment of VWB DNA containing and cloned in a non-replicative vector carrying a thiostrepton-resistance gene. Integration of pKT02 was obtained after transformation of ETH14630 and TK24 protoplasts. This vector will thus be useful for a number of additional species in which a suitable tRNA gene can be functional as integration site.


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