Transformation of Lactobacillus casei by electroporation

Chassy, Bruce M.

doi:10.1016/0378-1097(87)90315-6

Cited by 74 publications

(101 citation statements)

References 11 publications

(15 reference statements)

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“…transformation of Lactobacillus acidophilus by chromosomal DNA (Lin and Savage, 1986) and plasmid transformation in Lactobacillus reuteri (Morelli et al, 1987) have been demonstrated. Electroporation can be used to introduce DNA into cells of Laclobacillus case; (Chassy and Flickinger, 1987) and is not dependent on cell competence whir-!l is lacking in Laclobacillus case; and other lactic acid bacteria (Kondo and McKay, 1985). This method also does not require preparation of protoplasts and the specialized medium necessary to regenerate them.…”

Section: Transformationmentioning

confidence: 99%

Proline-Specific Peptidases of Lactobacillus casei Subspecies

Najafi

Lee

1994

Journal of Dairy Science

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Section: Transformationmentioning

confidence: 99%

Proline-Specific Peptidases of Lactobacillus casei Subspecies

Najafi

Lee

1994

Journal of Dairy Science

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“…Electroporation is also applied to the transformation of bacterial microorganisms. First example was the stimulation of transformation of Bacillus cereus protoplasts by electric field pulses (Shivarova et al 1983); more recently the electroporation technique was successfully applied to transform Streptococcus thermophilus (Somkuti and Steinberg 1987), Streptomyces lividans protoplast (Mac Nell 1987), Lactobacillus casei (Chassy and Flickinger 1987), Campylobacterjejuni (Miller et al 1988), different strains of Lactic streptococci (Powell et al 1988) Enterococcus faecalis protoplasts, Pseudomonas putida and Escherichia coli (Fiedler and Wirth;Taketo 1988). Although Corynebacteria osmotically sensitive cells can be transformed by the PEG technique, there is actually no method available to transform intact cells.…”

Section: Introductionmentioning

confidence: 99%

Electrotransformation of intact and osmotically sensitive cells of Corynebacterium glutamicum

Wolf

Pühler

Neumann

1989

Appl Microbiol Biotechnol

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Summary. Intact and osmotically sensitive cells ofCorynebacterium glutamicum can be efficiently transformed by electroporation. This was shown by using the plasmid vector pUL-330 (5.2 kb), containing the kanamycin resistance gene of transposon Tn5. The following electric parameters yielded efficient transformation. For intact cells: one exponentially decaying field pulse E = Eo exp ( -t/re) with time constants re = 450-500 ~ts and with initial field intensities of Eo= 35-40 kV cm-l; prepulse temperature 20 ° C. Cell regeneration (survival) was 100%-80%. Transformation efficiency can be increased by an additional freeze and thaw cycle of the cells, prior to electropotation. Lysozyme treated cells (osmotically sensitive) were transformed with three successive pulses of Eo = 25-30 kV cm-1. Cell regeneration under these conditions was found to be 20-30%. The optimum yield of transformants/~tg plasmid-DNA was 3 × 103 for intact cells, 2 × 104 for intact cells which were frozen and thawed twice and 7 x 104 for osmotically sensitive cells if the cell suspension was pulsed at a cell density of 1-3 x 108/ml and at a DNA concentration of 0.2 Ixg/ ml up to _< 2 Ixg/ml. The data obtained for osmotically sensitive cells suggest that the temperature increase accompanying the electric field pulse enhances colony formation and transformation efficiency if the initial prepulse temperature is _> 20 ° C, although regeneration of electroporated C. 91utamicum cells starts to decrease at temperatures > 20 ° C.

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“…lactis (42), L. plantarum (7), L. pentosus (29), and L. reuteri (16). Electroporation procedures have been developed for a number of Lactobacillus species, including L. acidophilus (41), L. casei (10,29), L. delbrueckii subsp. lactis (42), and L. helveticus (5,11).…”

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Comparative Sequence Analysis of Plasmids from Lactobacillus delbrueckii and Construction of a Shuttle Cloning Vector

Lee

Halgerson

Kim

et al. 2007

Appl Environ Microbiol

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While plasmids are very commonly associated with the majority of the lactic acid bacteria, they are only very rarely associated with Lactobacillus delbrueckii, with only four characterized to date. In this study, the complete sequence of a native plasmid, pDOJ1, from a strain of Lactobacillus delbrueckii subsp. bulgaricus was determined. It consisted of a circular DNA molecule of 6,220 bp with a G؉C content of 44.6% and a characteristic ori and encoded six open reading frames (ORFs), of which functions could be predicted for three-a mobilization (Mob) protein, a transposase, and a fused primase-helicase replication protein. Comparative analysis of pDOJ1 and the other available L. delbrueckii plasmids (pLBB1, pJBL2, pN42, and pLL1212) revealed a very similar organization and amino acid identities between 85 and 98% for the putative proteins of all six predicted ORFs from pDOJ1, reflecting a common origin for L. delbrueckii plasmids. Analysis of the fused primasehelicase replication gene found a similar fused organization only in the theta replicating group B plasmids from Streptococcus thermophilus. This observation and the ability of the replicon to function in S. thermophilus support the idea that the origin of plasmids in L. delbrueckii was likely from S. thermophilus. This may reflect the close association of these two species in dairy fermentations, particularly yogurt production. As no vector based on plasmid replicons from L. delbrueckii has previously been constructed, an Escherichia coli-L. delbrueckii shuttle cloning vector, pDOJ4, was constructed from pDOJ1, the p15A ori, the chloramphenicol resistance gene of pCI372, and the lacZ polylinker from pUC18. This cloning vector was successfully introduced into E. coli, L. delbrueckii subsp. bulgaricus, S. thermophilus, and Lactococcus lactis. This shuttle cloning vector provides a new tool for molecular analysis of Lactobacillus delbrueckii and other lactic acid bacteria.Bacteria of the genus Lactobacillus are gram-positive rods and constitute a major genus of the lactic acid bacteria, a group of economically important microorganisms that produce lactic acid as a major metabolite and are used in many fermentation processes as well as for probiotic purposes (20,25,32). Lactobacillus delbrueckii subsp. bulgaricus is used extensively in food fermentations, but especially in the production of yogurt. The organism was the original probiotic culture proposed by Metchnikoff (22), but its limited acid tolerance subsequently decreased its probiotic relevance. While the complete genome sequence of this bacterium has recently been published (18) Unlike the majority of Lactobacillus species, plasmids are extremely rare for L. delbrueckii (21). In a study of 48 strains of L. delbrueckii subsp. bulgaricus, a small cryptic plasmid was observed for only one strain, suggesting a frequency of Ͻ2% for these bacteria (3). To date, only one L. delbrueckii subsp. bulgaricus plasmid, pLBB1, has been isolated and sequenced (3). The plasmids pJBL2 and pN42 (6) and pLL1212 (GenBank ...

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Transformation of Lactobacillus casei by electroporation

Cited by 74 publications

References 11 publications

Proline-Specific Peptidases of Lactobacillus casei Subspecies

Proline-Specific Peptidases of Lactobacillus casei Subspecies

Electrotransformation of intact and osmotically sensitive cells of Corynebacterium glutamicum

Comparative Sequence Analysis of Plasmids from Lactobacillus delbrueckii and Construction of a Shuttle Cloning Vector

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