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Constitutive Gene Expression System for Lactococcus lactis and Other Lactic Acid Bacteria

ELV01200 - NICE pNZ2103 Lactococcus lactis const. vector, lacA; 10ug

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Constitutive Gene Expression System for 
Lactococcus lactis and Other Lactic Acid Bacteria

Features
  • Less endogenous and no exogenous proteases
  • Endotoxin-free expression system
  • Food grade protein expression possible
  • No formation of inclusion bodies
  • No formation of spores
  • Simple fermentation, scale-up, and downstream processing 
  • Suitable also for bacteria other than Lactococcus lactis
Areas of Application
  • Overexpression of homologous and heterologous genes for functional studies and to obtain large quantities of specific gene products
  • Metabolic engineering
  • Suitability for protein secretion and anchoring in the cell envelope
  • Large scale applications
Next to the wealth of traditional food applicatons, L. lactis is increasingly used for modern biotechnological applications such as the production of recombinant proteins for food, feed, pharmacological and biocatalysis applications. 

Description of pNZ2103 - pNZ2103 - Broad host range vector for constitutive gene expression under control of the lacA promoter. For transcriptional fusion. 

Plasmids

The replicons of the vectors pNZ124, pNZ2105, pNZ2103, pNZ7021, pNZ2122, and pNZ2123 are identical and came originally from the Lactococcus lactis plasmid pSH71. However, this replicon has a broad host range. Plasmids with this replicon can replicate in many Gram-positive bacteria, such as Lactobacillus plantarum and Streptococcus thermophilus. 

  • pNZ124 – General broad host range cloning vector with multiple cloning site for L. lactis and other lactic acid bacteria. Chloramphenicol selection. (Platteeuw et al., 1994)
  • pNZ2105 – General food grade cloning vector with multiple cloning site for L. lactis NZ3000LacF-based selection. (Platteeuw et al., 1996)
  • pNZ2103 – Broad host range vector for constitutive gene expression under control of the lacA promoter. For transcriptional fusion. Chloramphenicol selection. (Platteeuw et al., 1996)
  • pNZ7021 – Broad host range vector for constitutive gene expression under control of the strong pepN promoter. For transcriptional fusion. Chloramphenicol selection. (Wegkamp et al., 2007)
  • pNZ2122 – Food grade vector for constitutive gene expression under control of the lacA promoter for L. lactis NZ3000. For transcriptional fusions. LacF-based selection. (Platteeuw et al., 1996)
  • pNZ2123 – Food grade vector. Identical to pNZ2122, but with inverted multiple cloning site (MCS). LacF-based selection. (Platteeuw et al., 1996)

Host Strains
Lactococcus lactis subsp. cremoris MG1363
Plasmid-free progeny of the dairy strain NCDO712. Most widely used host strain for cloning and gene expression in L. lactis (Gasson, 1983).

Lactococcus lactis NZ3000
Standard strain for food grade selection based upon the ability to grow on lactose. The lactose operon, that is generally present on plasmids, has been integrated into the chromosome and the lacF gene was deleted. Delection of the lacF gene makes this strain unable to grow on lactose unless lacF is provided on a plasmid (van Alen-Boerrigter, I.J., and W.M. de Vos, unpublished data; de Ruyter et al., 1996).