pTCF176 HLA-A*02:01 (CD8 mut)
(Plasmid #180459)

• Purpose: E. coli expression of soluble MHC protein
• Depositing Lab: John Altman
• Publication: Altman lab bacterial expression plasmids (unpublished)

Backbone

• Vector backbone: pET24a+
• Backbone manufacturer: Novagen
• Vector type: Bacterial Expression

Growth in Bacteria

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

Gene/Insert

• Gene/Insert name: HLA-A*02:01 (CD8 mut)
• Species: H. sapiens (human)
• Mutation: D227K, T228A, replacement of transmembrane domain with enzymatic biotinylation site
• Tag / Fusion Protein:
  • BSP41 (C terminal on insert)

Cloning Information

• Cloning method: Restriction Enzyme
• 5′ cloning site: Unknown (unknown if destroyed)

Resource Information

• Supplemental Documents:
  • 180459: pTCF176 HLA-A*02:01 (CD8 mut)
  • pTCF176 HLA-A0201 (CD8 mut).gbk
• A portion of this plasmid was derived from a plasmid made by: synthetic cDNA, E. coli optimized

Terms and Licenses

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

Depositor Comments

Mutation prevents binding of CD8 (Purbhoo et al. (2001) J. Biol. Chem. doi.org/10.1074/jbc.M102498200) See https://www.ebi.ac.uk/ipd/imgt/hla/alleles/allele/?accession=HLA00005 for allele sequence data. For detailed protocols of expression, purification and use to visualize antigen-specific T cells, please refer to Altman and Davis (2016) Curr. Protoc. Immunol. doi.org/10.1002/cpim.14. We recommend the E. coli BL21(DE3) strain for expression.