Open AccessCloning and sequence determination of the gene for the human immunoglobulin epsilon chain expressed in a myeloma cell line.
Author(s) -
John H. Kenten,
H. V. Molgaard,
Michael Houghton,
R. Derbyshire,
Joanne L. Viney,
Lynda Bell,
Hannah J. Gould
Publication year - 1982
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.79.21.6661
Subject(s) - microbiology and biotechnology , complementary dna , biology , peptide sequence , oligonucleotide , nucleic acid sequence , gene , signal peptide , dna , molecular cloning , restriction enzyme , messenger rna , sequence analysis , immunoglobulin light chain , genetics , antibody
Messenger RNA has been isolated from cells of the human myeloma line 266BL which synthesizes IgE of the myeloma ND. A fraction enriched in mRNA for the epsilon heavy chain was copied into DNA and the DNA was cloned in Escherichia coli. A chemically synthesized oligonucleotide probe, based on the experimentally determined sequence of the specific message, was used to screen colonies. The largest epsilon chain cDNA cloned, 2.0 kilobases, was characterized by restriction endonuclease mapping and DNA sequence analysis. It appears to encode the complete amino acid sequence of the epsilon chain, including a signal peptide at the NH2 terminus as well as untranslated sequences at the 5' and 3' ends of the mRNA. The missing part of the previously published amino acid sequence of the ND epsilon chain was determined from the DNA sequence.