|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||46888||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size (bp) 8539
Modifications to backboneSee associated article for detailed construction information
Vector typeBacterial Expression ; Nisin-inducible expression; Gram-positive Bacterial Shuttle Vector
- Promoter PnisA
Growth in Bacteria
Growth instructionsFor 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 numberLow Copy
- Cloning method Restriction Enzyme
- 5′ sequencing primer Unknown
- 3′ sequencing primer T7; pBR322ori-F (5'-GGGAAACGCCTGGTATCTTT-3') (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byThe pNisA promoter and the nisin-sensing genes come from the Netherlands Dairy Research Institute (NIZO) (de Ruyter, Kuipers, and de Vos, 1996. PMID: 8837421)
Terms and Licenses
- Not Available to Industry
Articles Citing this Plasmid
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.
These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
For your Materials & Methods section:pMSP3545 was a gift from Gary Dunny (Addgene plasmid # 46888 ; http://n2t.net/addgene:46888 ; RRID:Addgene_46888)
For your References section:Improved vectors for nisin-controlled expression in gram-positive bacteria. Bryan EM, Bae T, Kleerebezem M, Dunny GM. Plasmid. 2000 Sep;44(2):183-90. 10.1006/plas.2000.1484 PubMed 10964628