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1 Department of Microbiology, Nash Hall 220, and The Center for Gene Research and Biotechnology, Oregon State University, Corvallis 97331-3804
2 Department of Microbiology, Nash Hall 220, and The Western Dairy Center, Utah State University, Logan 84322
The objective of this study was to construct a food-grade, phage-resistant strain of Lactococcus lactis by replacing a specific chromosomal gene with an allele that had been mutated in vitro. Lactococcus lactis contains a chromosomal gene (pip) that is required for infection by bacteriophages of the c2 species. A nonsense mutation in pip was constructed in vitro. The wild-type pip on the chromosome of strain LM2301 was exchanged for the mutated pip. The exchange left no antibiotic resistance genes or nonlactococcal DNA in the engineered strain (JK101). JK101 was resistant to the same phages as a strain that contains a spontaneous mutation in pip. JK101 grew as well as the pip+ isogenic strain did in minimal or rich media.
Key Words: phage resistance • bacteriophage • pip • gene replacement
Submitted on October 7, 1996
Accepted on February 18, 1997
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