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Nonsense suppressor strains of Lactococcus lactis were isolated using plasmids containing nonsense mutations or as revertants of a nonsense auxotrophic mutant. The nonsense suppressor gene was cloned from two suppressor strains and the DNA sequence determined. One suppressor is an ochre suppressor with an altered tRNA(gln) and the other an amber suppressor with an altered tRNA(ser). The nonsense suppressors allowed isolation of nonsense mutants of a lytic bacteriophage and suppressible auxotrophic mutants of L. lactis MG1363. A food-grade cloning vector based totally on DNA from Lactococcus and a synthetic polylinker with 11 unique restriction sites was constructed using the ochre suppressor as a selectable marker. Selection, following electroporation of a suppressible purine auxotroph, can be done on purine-free medium. The pepN gene from L. lactis Wg2 was subcloned resulting in a food-grade plasmid giving a four- to fivefold increase in lysine aminopeptidase activity.
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F Dickely, and
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E Johansen
April 2011,
Biotechnology letters,
F Dickely, and
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March 1995,
FEMS microbiology letters,
F Dickely, and
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April 2000,
Applied and environmental microbiology,
F Dickely, and
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June 2013,
Molecular biotechnology,
F Dickely, and
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January 2005,
Journal of applied microbiology,
F Dickely, and
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March 2008,
Wei sheng wu xue bao = Acta microbiologica Sinica,
F Dickely, and
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April 1998,
Applied microbiology and biotechnology,
F Dickely, and
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E B Hansen, and
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February 2011,
Current microbiology,
F Dickely, and
D Nilsson, and
E B Hansen, and
E Johansen
August 2022,
Folia microbiologica,
