Production of multiple murine CD2 receptor constructs using the baculovirus expression vector and a rapid dot-blot assay.
The baculovirus expression system was used to produce three different constructs of the murine cell surface adhesion receptor CD2. One construct coded for a single, N-terminal, Ig-fold domain. It was inefficiently secreted and therefore primarily intracellular. The second construct coded for both extracellular, N-terminal Ig-fold domains. This was efficiently secreted into culture supernatant. The third construct coded for the full-length transmembrane molecule which localized to the cell surface. All constructs were monomers of predicted MWr and were appropriately glycosylated. They retained epitopic specificity as demonstrated by binding to mAbs, and adhesion function as demonstrated by a rosetting assay.
Kaplan
A J
AJ
Department of Surgery, Medical University of South Carolina, Charleston.
Chavin
K D
KD
Qin
L H
LH
Yagita
H
H
Bromberg
J S
JS
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D016367:Baculoviridae; D018801:CD2 Antigens / Q000096:biosynthesis* / Q000276:immunology; D002460:Cell Line; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D005822:Genetic Vectors; D015151:Immunoblotting / Q000379:methods; D011994:Recombinant Proteins / Q000096:biosynthesis*; D018411:Spodoptera
D000911:Antibodies, Monoclonal; D018801:CD2 Antigens; D011994:Recombinant Proteins