GgVcl 1-1066 FRET tail probe 1
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||46276||Standard format: Plasmid sent in bacteria as agar stab||1||$75|
This material is available to academics and nonprofits only.
Backbone manufacturerCraig lab
Modifications to backbonepECFP-N3 vector was constructed as an intermediate vector for generating tail probe. ECFP was PCR amplified with 5-GGTACCatggtgagcaagggc-3 and 5-GCGGCCGCTttacttgtacagctc-3 to generate 5' KpnI and 3' NotI. The product was subcloned into TOPO pCRII and sequenced. The KpnI and NotI fragment of pCRII/ECFP was used to replace the EGFP fragment of pEGFP-N3 to generate pECFP-N3.
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Gene/Insert nameVcl 1-1066 + EYFP
SpeciesG. gallus (chicken)
Mutationaddition of 9 bp linker and EYFP at aa 883
- Promoter CMV
/ Fusion Protein
- ECFP (C terminal on backbone)
- Cloning method Restriction Enzyme
- 5′ cloning site EcoRI (not destroyed)
- 3′ cloning site SalI (not destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer ? (Common Sequencing Primers)
Terms and Licenses
- Not Available to Industry
A 9-bp fragment encoding a NotI site was introduced by mutagenesis into pEGFP-C1/vinculin cDNA (chicken) immediately after the codon for aa 883 (Coutu and Craig, 1988) using
the Quick Change kit (Stratagene). The cDNA of EYFP (CLONTECH Laboratories, Inc.) minus the stop codon and flanked by NotI was inserted into NotI-digested vinculin.
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:GgVcl 1-1066 FRET tail probe 1 was a gift from Susan Craig (Addgene plasmid # 46276 ; http://n2t.net/addgene:46276 ; RRID:Addgene_46276)
For your References section:Spatial distribution and functional significance of activated vinculin in living cells. Chen H, Cohen DM, Choudhury DM, Kioka N, Craig SW. J Cell Biol. 2005 May 9;169(3):459-70. 10.1083/jcb.200410100 PubMed 15883197