Open AccessExpression of a biologically active fragment of human IgE epsilon chain in Escherichia coli.
Author(s) -
FuTong Liu,
Keith Albrandt,
Cheryl G. Bry,
Teruko Ishizaka
Publication year - 1984
Publication title -
proceedings of the national academy of sciences of the united states of america
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.011
H-Index - 771
eISSN - 1091-6490
pISSN - 0027-8424
DOI - 10.1073/pnas.81.17.5369
Subject(s) - complementary dna , microbiology and biotechnology , escherichia coli , biology , expression vector , immunoglobulin e , coding region , untranslated region , northern blot , nucleic acid sequence , recombinant dna , messenger rna , gene , biochemistry , antibody , genetics
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.