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Related Experiment Videos

Food-grade delivery system for controlled gene expression in Lactococcus lactis.

B Henrich1, J R Klein, B Weber

  • 1Fachbereich Biologie, Abteilung Mikrobiologie, Universität Kaiserslautern, D-67653 Kaiserslautern, Germany. henrich@rhrk.uni-kl.de

Applied and Environmental Microbiology
|October 31, 2002
PubMed
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A novel food-grade system enables gene delivery and inducible expression in Lactococcus lactis. This system facilitates gene transfer to related strains, enhancing genetic engineering capabilities for dairy applications.

Area of Science:

  • Microbiology
  • Molecular Biology
  • Food Biotechnology

Background:

  • Lactococcus lactis is a key bacterium in dairy fermentation.
  • Efficient genetic manipulation systems are crucial for improving L. lactis strains.
  • Existing methods may lack food-grade compliance or efficient gene transfer.

Purpose of the Study:

  • To establish a food-grade system for gene delivery, inducible expression, and transfer in Lactococcus lactis.
  • To integrate and express heterologous genes, specifically peptidase genes, in L. lactis.
  • To demonstrate the transferability of engineered genetic elements between L. lactis strains.

Main Methods:

  • Construction of thermosensitive plasmid vectors based on the pG(+)host replicon.
  • Integration of genes into the L. lactis chromosome or sex factor via single crossovers.

Related Experiment Videos

  • Utilized the nisin-controlled expression (NICE) system for inducible gene transcription.
  • Conjugation was used to transfer engineered sex factors between L. lactis strains.

    • Main Results:

    • Successfully constructed food-grade L. lactis recombinants with integrated signal transduction genes (nisRK) and peptidase (pep) gene fusions.
    • Demonstrated inducible expression of P(nisA)::pep fusions using the NICE system after nisin induction.
    • Observed variable induction rates depending on the integration locus of nisRK.
    • Achieved successful transfer of an engineered sex factor carrying a P(nisA)::pepI fusion via conjugation at a frequency of 4 x 10(-4).

    Conclusions:

    • A versatile food-grade genetic system for L. lactis has been developed.
    • The system allows for inducible expression of cloned genes and efficient gene transfer.
    • This platform holds potential for metabolic engineering of L. lactis for improved food production.