Cloning and characterization of the determinant for abortive infection of bacteriophage from lactococcal plasmid pCI829

Abstract

The genetic determinant for abortive infection of bacteriophage (Abi) from the lactococcal plasmid pCI829 was cloned on a 6.2 kb StuI fragment in Escherichia coli using the shuttle vector pSA3. In Lactococcus lactis subsp. lactis MG1363Sm the resulting recombinant plasmid pCI816 conferred complete insensitivity to the small isometric-headed phage 712 and a reduced plaque size in the case of the prolate-headed phage c2. The determinant was further localized by subcloning and nuclease Bal31 deletion analysis; approximately 2.0 kb of DNA was essential for the expression of the Abi+ phenotype. Nucleotide sequence analysis of this region revealed a putative open reading frame of 1887 base pairs preceded by a putative promotor sequence and ribosome-binding site which exhibited similarity to consensus E. coli and Bacillus subtilis transcription/translation signals. Hybridization experiments indicated that this region was not homologous to the abi determinant from the phenotypically similar lactococcal plasmid pCI750.