Fusion constructs were cloned through overlap extension PCR (32 (link)) using wild type human CD137 (UniProt: Q07011; NCBI ref: NM_001561), human CD52 (Uniprot: P31358; NCBI ref: NM_001803.2) or the Rp3/Cp11 epitope (kind gift of Prof Martin Pule and Dr Brian Philip, UCL (33 (link), 34 (link))). Each fusion protein consisted of the CD52 leader sequence, Rp3/Cp11, CD137 (with domains removed based on the annotations provided with Uniprot reference sequence). These gene constructs were cloned into pcDNA3.1/- (neo) expression vector (Invitrogen).
Protocol detail
Engineered Fusion Proteins for Cell Targeting
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Cd137
Epitope
Gene
Human
Leader sequence
Protein
Vector
Corresponding Organization : University of Southampton
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Variable analysis
independent variables
- Fusion constructs cloned through overlap extension PCR
- Wild type human CD137 (UniProt: Q07011; NCBI ref: NM_001561)
- Human CD52 (Uniprot: P31358; NCBI ref: NM_001803.2)
- Rp3/Cp11 epitope (kind gift of Prof Martin Pule and Dr Brian Philip, UCL)
- Not explicitly mentioned
- CD52 leader sequence
- Removal of domains from CD137 based on Uniprot reference sequence annotations
- Cloning of gene constructs into pcDNA3.1/- (neo) expression vector (Invitrogen)
- Not explicitly mentioned
- Not explicitly mentioned
Annotations
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