Cloning of Mouse Immunoglobulin ε Gene and Its Location within the Heavy Chain Gene Cluster

• Published in: Proceedings of the National Academy of Sciences - PNAS 1981-03, Vol.78 (3), p.1581-1585
• Main Authors: Nishida, Yasuyoshi, Kataoka, Tohru, Ishida, Norio, Nakai, Sumiko, Kishimoto, Tadamitsu, Böttcher, Irmgard, Honjo, Tasuku
• Format: Article
• Language: English
• Subjects: Amino acids; Animals; Base Sequence; Cell Line; Cloning, Molecular; Complementary DNA; DNA; DNA probes; DNA Restriction Enzymes; DNA, Recombinant - metabolism; Gels; Genes; Genetic hybridization; Genetic screening; Humans; Hybrid Cells - immunology; Hybridomas; Immunoglobulin E - genetics; Immunoglobulin epsilon-Chains - genetics; Immunoglobulin G - genetics; Immunoglobulin Heavy Chains - genetics; Mice; Mice, Inbred BALB C; Nucleic Acid Hybridization; Nucleotide sequences; Plasmacytoma; Spleen - immunology
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.78.3.1581

Mouse immunoglobulin ε chain gene was cloned from DNA of a hybridoma producing anti-dinitrophenyl IgE, which was constructed by fusing a spleen cell of a BALB/c mouse with a variant clone of MOPC21 myeloma (IgG1 producer). Because a given active heavy chain constant region (CH) gene is linked to a heavy chain joining segment (JH) gene at its 5′side, the expressed Cεgene of the hybridoma was cloned from a phage library containing partial Sau3A digests of IgE hybridoma DNA by using a J gene fragment as a probe. Among 6× 105phages screened, five positive clones were obtained and three of them were identified as Cεgene clones by restriction mapping, Southern blot hybridization, R-loop formation, and partial nucleotide sequence determination. The determined nucleotide sequence predicted the amino acid sequence which resembles a part of the CH3domain of human ε chain. The deletion profile of the Cεgene in various myelomas expressing different CHgenes indicates that the Cεgene is located between the Cγ 2aand Cαgenes. The linkage (5′-ε -α -3′) was directly confirmed by molecular cloning of the overlapping chromosomal segments from newborn mouse DNA.