|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||22007||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) 6000
Vector typeMammalian Expression, Bacterial Expression, Insect Expression
Growth in Bacteria
Copy numberHigh Copy
SpeciesH. sapiens (human); Avena sativa (oat)
Insert Size (bp)1100
MutationRac1 starts at I4 and contains mutations Q61L, E91H and N92H.
Entrez GeneRAC1 (a.k.a. MIG5, MRD48, Rac-1, TC-25, p21-Rac1)
/ Fusion Proteins
- 6X His (N terminal on backbone)
- mVenus (N terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site Bam HI (not destroyed)
- 3′ cloning site HindIII (not destroyed)
- 5′ sequencing primer pTriExUP (Common Sequencing Primers)
Protocols for production and use of Hahn lab biosensors and photoactivatable proteins can be found on their web page, at the URL below. If you have suggestions for the protocols or have any questions, please feel free to contact the Hahn lab. Good luck with your experiments!
Hahn lab biosensor protocol: http://www.hahnlab.com/tools/index.html
Please note, Addgene has not confirmed all the mutations associated with this construct. We suggest sequence verifying this plasmid upon receipt.
Further remarks from the Hahn lab regarding some mutations in the fluorescent marker: "the C-S mutation was introduced by us to allow site specific labeling of the protein and we have not detected any effect.
The F-L mutation reverts the Venus sequence back to the YFP sequence at this site. It will have a very slight detrimental effect on the maturation speed of the chromophore, but will not affect the stability or wavelength of the chromophore.
Neither of these 2 mutations will have any effect on the function of the Lov-Rac construct , and the Venus will still function as it should, i.e. as a reporter for the 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:pTriEx-mVenus-PA-Rac1 was a gift from Klaus Hahn (Addgene plasmid # 22007 ; http://n2t.net/addgene:22007 ; RRID:Addgene_22007)
For your References section:A genetically encoded photoactivatable Rac controls the motility of living cells. Wu YI, Frey D, Lungu OI, Jaehrig A, Schlichting I, Kuhlman B, Hahn KM. Nature. 2009 Sep 3. 461(7260):104-8. 10.1038/nature08241 PubMed 19693014