Resource Summary Report  New Search Previous Search Results

URL: http://www.addgene.org/69600

Proper Citation: RRID:Addgene_69600

Bacterial Resistance: Ampicillin

Defining Citation: PMID:9553774

Vector Backbone Description: Backbone Marker:Invitrogen; Vector Backbone:pBluescript KS; Vector Types:Mammalian Expression; Bacterial Resistance:Ampicillin

Comments: 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.

Expand All Collapse