pF142 pAcGFP1 SOD1 A4V
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||26403||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 (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer SV40pA-R (Common Sequencing Primers)
A portion of this plasmid was derived from a plasmid made byhuman cDNA insert from cDNA clone cSOD1 from Professor Yoram Groner as described in Lieman-Hurwitzet al., 1982 and Sherman et al 1983.
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 4 of the cDNA from GCC to GTC, resulting in the A4V 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 pACGFPC1. Thus GFP is 5’ of the SOD1 insert and upstream in the GFP-SOD1 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:pF142 pAcGFP1 SOD1 A4V was a gift from Elizabeth Fisher (Addgene plasmid # 26403 ; http://n2t.net/addgene:26403 ; RRID:Addgene_26403)
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