Transformation of Lactococcus by Electroporation
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Last Update: 2021-02-22
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Source: Internet
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Author: User
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Lactococcus lactis
strains are the principal organisms used in cheese-making, and their performance is of vital importance to the quality of the cheese. A goal for the dairy industry has always been to improve strains of
L. lactis
and to stabilize beneficial traits. In recent years, genetic engineering has become a useful tool for manipulating
L. lactis
. A critical step in this work is the introduction of
DNA
into the cells. The first successful transformations of
L. lactis
employed protoplast transformation in the presence of polyethylene glycol (
1
). This method was not very reproducible and worked only for a few strains. More recently, electroporation proved to be an alternative to protoplast transformation in
L. lactis
. Harlander (
2
) was the first to employ electroporation in this species. Later work indicated that the cell wall is a physical barrier to the entering DNA not only during protoplast transformation, but also during electroporation. Powell et al. (
3
) used lysozyme treatment to obtain an increase in transformation efficiency, and van der Lelie et al. (
4
) obtained cells competent for electrotransformation by growing them in the presence of high concentrations of threonine to weaken their cell walls. However, the methods were not very efficient, and a number of strains could not be transformed.
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