Abstract
The 6.5 kb HindIII DNA fragment of the Lactococcus lactis subsp. cremoris H2 plasmid pDI21 was cloned into Escherichia coli POP13 with λNM1149, and also directly into Lactococcus lactis subsp. lactis 4125 using a newly-constructed broad host-range vector pFX1. Proteinase was experessed in both transformed organisms. The proteinase resembles a PI type since it preferentially degraded β-casein. The restriction map of the 6.5 kb proteinase gene fragment has minor differences from those of published plamid proteinase genes. High-efficiency electroporation with pFX1 provides a direct approach for gene cloning in lactococci.
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Abbreviations
- cfu:
-
colony forming units
- HEPES:
-
N-[2-hydroxyethyl]piperazine-N′-[2-ethanesulphonic acid]
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Dedicated to Prof. Dr. G. Drews on the occasion of his 65th birthday
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Xu, FF., Pearce, L.E. & Yu, PL. Molecular cloning and expression of a proteinase gene from Lactococcus lactis subsp. cremoris H2 and construction of a new lactococcal vector pFX1. Arch. Microbiol. 154, 99–104 (1990). https://doi.org/10.1007/BF00249185
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DOI: https://doi.org/10.1007/BF00249185