Incompatibility of
            <i>Lactobacillus</i>
            Vectors with Replicons Derived from Small Cryptic
            <i>Lactobacillus</i>
            Plasmids and Segregational Instability of the Introduced Vectors

Posno, M.; Leer, R.J.; Luijk, N. van; Giezen, M. J. F. van; Heuvelmans, P. T. H. M.; Lokman, B. Christien; Pouwels, Peter H.

doi:10.1128/aem.57.6.1822-1828.1991

Cited by 179 publications

(58 citation statements)

References 45 publications

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“…Plasmids isolated from L. lactis were used for electroporation of all Lactobacillus strains. Electroporation of L. casei was carried out as previously described [31]. The same protocol was used for the other lactobacillus strains except that the electroporation buffer was three-fold concentrated.…”

Section: Plasmids and Recombinant Dna Proceduresmentioning

confidence: 99%

Induction of systemic and mucosal immune response and decrease in Streptococcus pneumoniae colonization by nasal inoculation of mice with recombinant lactic acid bacteria expressing pneumococcal surface antigen A

Oliveira

Arêas

Campos

et al. 2006

Microbes and Infection

102

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Mucosal epithelia constitute the first barriers to be overcome by pathogens during infection. The induction of protective IgA in this location is important for the prevention of infection and can be achieved through different mucosal immunization strategies. Lactic acid bacteria have been tested in the last few years as live vectors for the delivery of antigens at mucosal sites, with promising results. In this work, Streptococcus pneumoniae PsaA antigen was expressed in different species of lactic acid bacteria, such as Lactococcus lactis, Lactobacillus casei, Lactobacillus plantarum, and Lactobacillus helveticus. After nasal inoculation of C57Bl/6 mice, their ability to induce both systemic (IgG in serum) and mucosal (IgA in saliva, nasal and bronchial washes) anti-PsaA antibodies was determined. Immunization with L. lactis MG1363 induced very low levels of IgA and IgG, possibly by the low amount of PsaA expressed in this strain and its short persistence in the nasal mucosa. All three lactobacilli persisted in the nasal mucosa for 3 days and produced a similar amount of PsaA protein (150-250 ng per 10(9) CFU). However, L. plantarum NCDO1193 and L. helveticus ATCC15009 elicited the highest antibody response (IgA and IgG). Vaccination with recombinant lactobacilli but not with recombinant L. lactis led to a decrease in S. pneumoniae recovery from nasal mucosa upon a colonization challenge. Our results confirm that certain Lactobacillus strains have intrinsic properties that make them suitable candidates for mucosal vaccination experiments.

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Section: Plasmids and Recombinant Dna Proceduresmentioning

confidence: 99%

Induction of systemic and mucosal immune response and decrease in Streptococcus pneumoniae colonization by nasal inoculation of mice with recombinant lactic acid bacteria expressing pneumococcal surface antigen A

Oliveira

Arêas

Campos

et al. 2006

Microbes and Infection

102

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“…Sm or Pm were used for counter-selection of the donor. The segregational stability of plasmids in di¡erent hosts was determined as described by Posno et al [14].…”

Section: Mating Proceduresmentioning

confidence: 99%

The broad host range plasmid pLF1311 from Lactobacillus fermentum VKM1311

Aleshin

1999

FEMS Microbiology Letters

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The complete nucleotide sequence (2389 bp) of the cryptic plasmid pLF1311 from Lactobacillus fermentum VKM1311 was determined. DNA sequence analysis revealed the putative coding regions for a replicative protein (RepB), its repressor (RepA) and double-stranded (dso) and single-stranded (sso) origins. pLF1311 belongs to the pE194 family of rolling circle-replicating plasmids. A derivative of pLF1311 that contains the cat gene of plasmid pC194 of Staphylococcus aureus and the oriT of RP4 was constructed and transferred by conjugative mobilization from Escherichia coli to various Gram-positive bacteria. The stable maintenance of this derivative was shown in some strains of Lactobacillus, Lactococcus, Enterococcus and Bacillus under non-selective conditions. ß

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“…Bioactive compounds must be expressed at a high enough level to elicit the desired immune response (3). While the high copy numbers of some plasmids may seem advantageous for antigen expression, plasmid instability and the selective pressure required for plasmid maintenance complicate their use in human clinical applications (13,14). Integrating genes into the chromosome for expression is expected to eliminate selection requirements and provide genetic stability (4).…”

mentioning

confidence: 99%

Chromosomal Insertions in the Lactobacillus casei upp Gene That Are Useful for Vaccine Expression

Song

et al. 2014

Appl Environ Microbiol

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bTo develop a stable and marker-free Lactobacillus strain useful for the expression of vaccines, we developed a temperature-sensitive suicide plasmid with expression cassettes containing an HCE promoter, a PgsA anchor, the alpha-toxin gene, and an rrnB T1T2 terminator (PP␣T) that uses a 5-fluorouracil (5-FU) counterselectable marker for Lactobacillus casei. Three strains containing the correct PP␣T expression cassettes were produced via the selective pressure of 5-FU screening. We confirmed that the upp gene was deleted and that the PP␣T expression cassettes were inserted into the upp site of L. casei ATCC 393 by genomic PCR amplification and sequencing. 5-FU resistance in recombinant bacteria could be stably inherited for as long as 40 generations following insertion. However, bacteria containing the integrated DNA grew more slowly than wild-type L. casei. An indirect enzyme-linked immunosorbent assay (ELISA) analysis demonstrated that the alpha-toxin gene was expressed. Also, we visualized expression of the protein on the surface of L. casei cells using laser confocal microscopy. These results taken together demonstrate that these recombinant bacteria should provide a safe tool for effective vaccine production.

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Incompatibility of Lactobacillus Vectors with Replicons Derived from Small Cryptic Lactobacillus Plasmids and Segregational Instability of the Introduced Vectors

Cited by 179 publications

References 45 publications

Induction of systemic and mucosal immune response and decrease in Streptococcus pneumoniae colonization by nasal inoculation of mice with recombinant lactic acid bacteria expressing pneumococcal surface antigen A

Induction of systemic and mucosal immune response and decrease in Streptococcus pneumoniae colonization by nasal inoculation of mice with recombinant lactic acid bacteria expressing pneumococcal surface antigen A

The broad host range plasmid pLF1311 from Lactobacillus fermentum VKM1311

Chromosomal Insertions in the Lactobacillus casei upp Gene That Are Useful for Vaccine Expression

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