pMSP3545
(Plasmid #46888)

• Purpose: (Empty Backbone) Gram-Positive Bacterial Shuttle Vector for Nisin-Controlled Inducible Expression
• Depositing Lab: Gary Dunny
• Publication: Bryan et al Plasmid. 2000 Sep;44(2):183-90.

Backbone

• Vector backbone: pMSP3545
• Backbone size (bp): 8539
• Modifications to backbone: See associated article for detailed construction information
• Vector type: Bacterial Expression ; Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector
• Promoter: PnisA

Growth in Bacteria

• Bacterial Resistance(s): Erythromycin, 200 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Growth instructions: For E. coli, grow on Luria Broth (LB) agar containing 300 μg/mL erythromycin or Brain Heart Infusion (BHI) medium containing 100 μg/mL erythromycin. For E. faecalis, grow on BHI containing 10 μg/mL erythromycin.
• Copy number: Low Copy

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ sequencing primer: Unknown
• 3′ sequencing primer: T7; pBR322ori-F (5'-GGGAAACGCCTGGTATCTTT-3')

Resource Information

• A portion of this plasmid was derived from a plasmid made by: The pNisA promoter and the nisin-sensing genes come from the Netherlands Dairy Research Institute (NIZO) (de Ruyter, Kuipers, and de Vos, 1996. PMID: 8837421)
• Articles Citing this Plasmid:
  • 5 References

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

This plasmid utilizes the NIsin Controlled Expression (NICE) system. NICE® is a registered trademark of NIZO food research BV, P.O.Box 20, 6710 BA Ede, The Netherlands.

pMSP3545 contains the Nisin-inducible PnisA promoter, and the pAMB1 replicon for expression in gram-positive bacteria. It also contains a unique NcoI cleavage site immediately downstream of the nisA ribosomal binding sequence, which may be used for translational fusions, and a rho-independent terminator between the XbaI and XhoI sites of the polylinker.

For protein expression in E. faecalis, induce with 10-25 ng/mL nisin (Sigma, N-5764). See associated article for instructions on how to make nisin solution.

Addgene's quality control sequencing finds discrepancies with the depositor's provided sequence, but they are not thought to affect plasmid function.