Subtractive cDNA Cloning
Dendritic cells (DC) represent a specific subset of leukocytes that play a critical role in the initiation of T, cell-mediated immune responses. Recently, we have established, from the epidermis of BALB/c mice, long-term DC lines, termed the “XS series”, which resemble resident epidermal DC (i.e., Langerhans cells) in many respects, including their morphology, surface phenotype, antigen presenting capacity, and cytokine and cytokine receptor mRNA profiles (l–3). Taking advantage of these stable lines, we sought to study unique attributes of DC at a molecular level by identifying molecules that they express selectively. For this aim, we employed a subtractive cDNA cloning strategy in which cDNA prepared from the XS52 DC line was subtracted with mRNA isolated from the i774 macrophage line. In this brief communication, we outline our methods as well as the identities of cDNAs isolated in this manner.
KeywordsSlot Blotting Colony Hybridization Macrophage Line J774 Macrophage Differential Hybridization
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