|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||52672||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
- Backbone size w/o insert (bp) 4733
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Alt namePostsynaptic density protein 95
Alt nameDisks large homolog 4 (DLG4)
Alt nameSynapse-associated protein 90 (SAP-90)
SpeciesR. norvegicus (rat)
Entrez GeneDlg4 (a.k.a. Dlgh4, PSD95, Sap90)
- Promoter CMV
/ Fusion Protein
- tagRFP (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site HindIII (not destroyed)
- 3′ cloning site EcoRI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer mKate2-TurboRFP-R (Common Sequencing Primers)
HindII and EcoRI were used to cut PSD-95 from PSD-95-pGW1 (received from David Bredt) and ligate it into pTagRFP-N. PSD-95 tyrosine 12 was mutated to glutamate (Y12E) by extending a complementary set of primers (PSD95Y12E-forward: 5’-GAC AAC CAA GAA AGA GCG CTA CCA AGA TGA AGA CAC G-3’, PSD95Y12E-reverse: 5'-CGT GTC TTC ATC TTG GTA GCG CTC TTT CTT GGT TGT C-3') along the whole template plasmid using Q5 high-fidelity DNA polymerase (NEB). DpnI was subsequently used to digest non-mutated, methylated template DNA prior to transformation.
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:PSD-95_Y12E-pTagRFP was a gift from Johannes Hell (Addgene plasmid # 52672 ; http://n2t.net/addgene:52672 ; RRID:Addgene_52672)