|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26409||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) 4700
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
SpeciesH. sapiens (human)
Insert Size (bp)470
Entrez GeneSOD1 (a.k.a. ALS, ALS1, HEL-S-44, IPOA, SOD, STAHP, hSod1, homodimer)
/ Fusion Protein
- GFP (C terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer CMV-F (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
The complete cDNA was subcloned into the vector pET28. Site directed mutagenesis changed wildtype DNA sequence for codon 37 of the cDNA from GGA to AGA, resulting in the G37R mutation in the protein and then a PCR amplified the complete cDNA with EcoRI and SalI sites on the 5’ and 3’ end respectively. This EcoRI-Sal1 0.47kb fragment was cloned into the respective EcoRI and SalI sites of pAcGFP1N1. Thus SOD1 is 5’ of GFP and SOD1 lies upstream of GFP in the SOD1-GFP fusion protein.
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:pF148 pSOD1G37RAcGFP1 was a gift from Elizabeth Fisher (Addgene plasmid # 26409 ; http://n2t.net/addgene:26409 ; RRID:Addgene_26409)
For your References section:Modification of superoxide dismutase 1 (SOD1) properties by a GFP tag--implications for research into amyotrophic lateral sclerosis (ALS). Stevens JC, Chia R, Hendriks WT, Bros-Facer V, van Minnen J, Martin JE, Jackson GS, Greensmith L, Schiavo G, Fisher EM. PLoS One. 2010 . 5(3):e9541. 10.1371/journal.pone.0009541 PubMed 20221404