ECFP-Bcr NE/AA
(Plasmid #36418)

• Depositing Labs: John Groffen, Nora Heisterkamp
• Publication: Cho et al Mol Cell Biol. 2007 Feb;27(3):899-911. Epub 2006 Nov 20.

Backbone

• Vector backbone: pECFP-C1
• Backbone manufacturer: Clontech
• Backbone size w/o insert (bp): 4700
• Total vector size (bp): 9000
• Vector type: Mammalian Expression
• Selectable markers: Neomycin (select with G418)

Growth in Bacteria

• Bacterial Resistance(s): Kanamycin, 50 μg/mL
• Growth Temperature: 37°C
• Growth Strain(s): DH5alpha
• Copy number: High Copy

Gene/Insert

• Gene/Insert name: breakpoint cluster region
• Alt name: BCR
• Species: H. sapiens (human)
• Insert Size (bp): 4300
• Mutation: N689A/ E690A
• Entrez Gene: BCR (a.k.a. ALL, BCR1, CML, D22S11, D22S662, PHL)
• Tag / Fusion Protein:
  • ECFP (N terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: BamHI (destroyed during cloning)
• 3′ cloning site: Kpn I (not destroyed)
• 5′ sequencing primer: EGFP-C
• 3′ sequencing primer: SV40pA-R

Resource Information

• Article Citing this Plasmid:
  • 1 Reference

Terms and Licenses

• Academic/Nonprofit Terms:
  • UBMTA
• Industry Terms:
  • Not Available to Industry

Depositor Comments

Made by mutagenesis directly on plasmid Bcr-ECFP using a Stratagene Quick Change kit. In Dbl, mutation of residues N673 and D674 completely abolished its activity as a GEF towards Rac. These residues are highly conserved among DH domains and are also conserved as residues N689/ E690 in Bcr. We therefore mutated these residues in Bcr and introduced the mutants into the RAW 264.7 macrophage cell line. However, the construct did not caused morphological changes in these cells. Constructed by Young-Jin Cho.