pTCF79 H-2D(b) (CD8 mut)
(Plasmid #180453)

• 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• Publication: Altman lab bacterial expression plasmids (unpublished)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: H-2D(b) (CD8 mut)
• Species: M. musculus (mouse)
• Mutation: D227K, replacement of transmembrane domain with enzymatic biotinylation site
• GenBank ID: NP_034510.3
• 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:
  • 180453: pTCF79 H-2D(b) (CD8 mut)
  • pTCF79 H-2D(b) (CD8 mut).gbk

Terms and Licenses

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

Depositor Comments

Mutation prevents binding of CD8 (Potter et al. (1989) Nature doi.org/10.1038/337073a0). 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.