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Cloning and Recombinant Protein Expression in Lactococcus lactis.
Susheel K Singh1, Mohammad Naghizadeh2,3, Jordan Plieskatt2
1Biotherapeutic and Vaccine Research Division, ICMR-Regional Medical Research Centre, Bhubaneswar, Odisha, India.
Methods in Molecular Biology (Clifton, N.J.)
|April 24, 2023
Summary
Lactococcus lactis is a safe and efficient bacterial host for producing functional recombinant proteins. This protocol details using L. lactis for high-level soluble protein expression and purification.
Area of Science:
- Microbiology
- Biotechnology
- Protein Expression
Background:
- Lactococcus lactis is a Gram-positive bacterium recognized as a safe and efficient host for recombinant protein production.
- Its desirable features include rapid growth, GRAS status, probiotic properties, and secretion of stable, properly folded proteins.
- Previous work demonstrated L. lactis with the P170 promoter for producing proteins of varying complexity.
Purpose of the Study:
- To provide a detailed protocol for utilizing L. lactis as a reliable platform for heterologous recombinant protein expression.
- To facilitate wider application of L. lactis for producing soluble recombinant proteins.
Main Methods:
- Cloning target genes into appropriate L. lactis expression vectors.
- Determining expression levels of the heterologous protein.
- Initial purification of expressed soluble recombinant proteins.
Main Results:
- Established a detailed protocol for L. lactis-based recombinant protein production.
- Demonstrated the feasibility of expressing and purifying soluble heterologous proteins.
Conclusions:
- Lactococcus lactis serves as a robust and versatile platform for the overproduction of functional recombinant proteins.
- The presented protocol enables broader adoption of L. lactis for soluble protein expression and purification.
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