Purpose(Empty Backbone) TCR beta chain transgene expression plasmid which has a unique ClaI site for the insertion of a TCR beta chain VDJ exon
Full plasmid sequence is not available for this item.
|Item||Catalog #||Description||Quantity||Price (USD)|
|Plasmid||69600||Standard format: Plasmid sent in bacteria as agar stab||1||$65|
This material is available to academics and nonprofits only.
Vector backbonepBluescript KS
Modifications to backboneSee comments
Vector typeMammalian Expression
Growth in Bacteria
- Cloning method Restriction Enzyme
Terms and Licenses
A 1.0 kb Sma-I/Not-I fragment containing the TCR b-chain enhancer element was excised from pKS913.Cb18.31 and cloned into the pKS
Bluescript plasmid (Stratagene), which had been modified to remove the Kpn-I and Cla-I restriction sites from the polylinker resulting in the pKS.bEnh plasmid. A Sal-I fragment, containing the B3 TCR b-chain enhancer, promoter and coding regions, was isolated from
pKS913.Cb18.31 and cloned into the Sal-I site of the pKS.bEnh
plasmid to give the construct pKS913.Cb18.31(Kpn-I)/Cla-I)). A
4.2 kb Eco-I/Cla-I fragment comprising the 5’ region and leader
sequence from the hybridoma 2B4 was isolated from the p3A9 b
shuttle vector. This fragment was cloned into the pGEM-T
plasmid (Promega) resulting in the pG3A9 plasmid. A
4.2 kb Apa-I/Cla-I fragment, containing the 2B4 5’ region and
leader sequence, was isolated from the pG3A9 plasmid. This
fragment was cloned into pKS319.Cb18.31(Kpn-I)/Cla-I)) after
removal of the B3 TCR b-chain promoter and the TCRBVDJ
coding regions by a combined Apa-I/Cla-I restriction enzyme digest, resulting in the p3A9cbTCR plasmid.
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:p3A9cbetaTCR was a gift from Francis R. Carbone (Addgene plasmid # 69600 ; http://n2t.net/addgene:69600 ; RRID:Addgene_69600)
For your References section:Defective TCR expression in transgenic mice constructed using cDNA-based alpha- and beta-chain genes under the control of heterologous regulatory elements. Barnden MJ, Allison J, Heath WR, Carbone FR. Immunol Cell Biol. 1998 Feb;76(1):34-40. 10.1046/j.1440-1711.1998.00709.x PubMed 9553774
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