Expression of a biologically active fragment of human IgE epsilon chain in Escherichia coli.
cDNA corresponding to human IgE heavy (epsilon) chain mRNA was cloned from human IgE-secreting myeloma U266 cells. Partial nucleotide sequence analysis demonstrated that the cloned cDNA contained the coding region for about two-thirds of the CH2 and all of the CH3 and CH4 domains as well as the 3'-untranslated region. This epsilon cDNA was inserted into expression vector pUC7 and expression of an epsilon-chain fragment in Escherichia coli was demonstrated by protein blot analysis using 125I-labeled goat anti-human IgE as probe. The expression product was purified on a column of goat anti-human IgE-conjugated Sepharose 4B and the polypeptide was found to retain binding activity to human basophils.
D000595:Amino Acid Sequence; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D004926:Escherichia coli / Q000235:genetics*; D006801:Humans; D007073:Immunoglobulin E / Q000235:genetics*; D007141:Immunoglobulin Fc Fragments / Q000235:genetics; D007143:Immunoglobulin Heavy Chains / Q000235:genetics*; D007139:Immunoglobulin epsilon-Chains / Q000235:genetics*; D009101:Multiple Myeloma / Q000276:immunology
D007141:Immunoglobulin Fc Fragments; D007143:Immunoglobulin Heavy Chains; D007139:Immunoglobulin epsilon-Chains; D007073:Immunoglobulin E; D004247:DNA; D004262:DNA Restriction Enzymes