pRc/CMV HA TrkB-T1
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||39980||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepRC/CMV AC7 vector, a derivative of the pRC/CMV vector
- Backbone size w/o insert (bp) 5500
Vector typeMammalian Expression
Selectable markersNeomycin (select with G418)
Growth in Bacteria
Copy numberHigh Copy
Alt nameNTrk2 variant 2
SpeciesR. norvegicus (rat)
Insert Size (bp)1500
MutationTrkB.T1 variant; endogenous signal sequence was replaced with BM40 and HA tag
Entrez GeneNtrk2 (a.k.a. RATTRKB1, TRKB1, Tkrb, trk-B, trkB)
- Promoter CMV
/ Fusion Proteins
- HA (N terminal on insert)
- BM40 signal sequence (N terminal on insert)
- Cloning method Restriction Enzyme
- 5′ cloning site NheI (unknown if destroyed)
- 3′ cloning site NotI (unknown if destroyed)
- 5′ sequencing primer CMV-F
- 3′ sequencing primer SP6 (Common Sequencing Primers)
The T1 isoform of trkB was cloned from a pCMV-5 trkB.T1 construct using HindIII and XbaI to fuse it to the HA-tagged N-terminus of trkB in the vector pB-KS followed by subcloning into pRC/CMV AC7 with NheI and NotI.
See Middlemas et al (Mol Cell Biol. 1991 Jan;11(1):143-53., https://www.ncbi.nlm.nih.gov/pubmed?term=1846020 ) to learn more about the TrkB.T1 variant.
Please note that the endogenous signal sequence of TrkB-T1 was replaced by the BM40 signal peptide sequence (MRAWIFFLLCLAGRALAAPLA). When compared to translated sequences for GenBank ID M55292.1 or NM_001163168.1, the TrkB-T1 protein sequence starts at amino acid #32
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:pRc/CMV HA TrkB-T1 was a gift from Yves-Alain Barde (Addgene plasmid # 39980 ; http://n2t.net/addgene:39980 ; RRID:Addgene_39980)
For your References section:Biochemical and functional interactions between the neurotrophin receptors trk and p75NTR. Bibel M, Hoppe E, Barde YA. EMBO J. 1999 Feb 1;18(3):616-22. 10.1093/emboj/18.3.616 PubMed 9927421