|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||36095||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Modifications to backboneTo create a yeast GFP expression vector (pGOGFP), the PRC1 promoter and GFP coding sequences were subcloned into the BamHI site of PRS426. To create a PtdIns(4)P-specific FLARE construct,pRS424GFP-2x(Osh2), we fused two tandem copies of the Osh2 PH domain to GFP. PCR was used to generate sequences encoding two Osh2 PH domains. These PCR products were cloned in-frame into pGOGFP (Burd and Emr, 1998), to create a plasmid harboring a GFP-2×PH(Osh2) fusion, pRS424GFP-2×PH(Osh2).
Vector typeYeast Expression
Growth in Bacteria
Gene/Insert name2xPH(Osh2) fusion
SpeciesS. cerevisiae (budding yeast)
- Promoter PRC1
/ Fusion Protein
- GFP (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site BamHI (destroyed during cloning)
- 3′ cloning site SalI (destroyed during cloning)
- 5′ sequencing primer T3
- 3′ sequencing primer T7 (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
Article Citing this Plasmid
Please note that the first Osh2 domain contains aa273-421 and the second Osh2 domains contains aa260-421 when compared to GenBank reference sequence NP_010265.1. According to this reference sequence the PH domain is aa 292–384. The additional residues surrounding the PH domain were intentionally cloned into this construct.
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:pRS424GFP-2xPH(Osh2) was a gift from Scott Emr (Addgene plasmid # 36095 ; http://n2t.net/addgene:36095 ; RRID:Addgene_36095)
For your References section:Osh proteins regulate phosphoinositide metabolism at ER-plasma membrane contact sites. Stefan CJ, Manford AG, Baird D, Yamada-Hanff J, Mao Y, Emr SD. Cell. 2011 Feb 4;144(3):389-401. 10.1016/j.cell.2010.12.034 PubMed 21295699
Map uploaded by the depositor.