Production of multiple murine CD2 receptor constructs using the baculovirus expression vector and a rapid dot-blot assay |
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Authors: | Adam J. Kaplan Kenneth D. Chavin Li Hui Qin Hideo Yagita Jonathan S. Bromberg |
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Affiliation: | 1. Department of Surgery, Medica University of South Carolina, Charleston, S.C., USA 2. Microbiology and Immunology, Medical University of South Carolina, Charleston, S.C., USA 3. Department of Immunology, Juntendo University, School of Medicine, Tokyo, Japan 4. Department of Surgery, University of Michigan, 2926 Taubman Center, 48109-0331, Ann Arbor, MI, USA
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Abstract: | 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. |
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