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PMID: 113775 Published · ppublish English Journal Article

Cloning of immunoglobulin kappa light chain genes from mouse liver and myeloma MOPC 173.

Nucleic acids research ·Vol. 6 ·No. 10 ·1979-07-25 ·Pages 3213-29

Steinmetz M, Zachau HG, Mach B

Abstract

The organization of the kappa chain constant region gene was compared in DNA from an immunoglobulin-producing mouse myeloma (MOPC 173) and from liver. In situ hybridization using the Southern blotting technique revealed constant region gene-containing EcoRI-DNA fragments of 14 and 20 kb in the myeloma tissue whereas one EcoRI-DNA fragment with a length of 15 kb was found in liver DNA. After enrichment by RPC-5 chromatography and preparative electrophoresis the 14 kb fragment from MOPC 173 DNA and the 15 kb fragment from liver DNA were cloned in the bacteriophage lambda vector Charon 4A using in vitro packaging. Extensive characterization of the two fragments by restriction endonuclease mapping, in situ hybridization, and electron microscopy (R-loop and heteroduplex) showed that both fragments contain the constant region but no MOPC 173 variable region gene. Both fragments are homologous over a length of 12.5 kb including the constant region but differ from one another starting about 2.7 kb from the 5' end of the constant region gene. This indicates that the 14 kb EcoRI-DNA fragment from the myeloma tissue clearly resulted from somatic DNA rearrangement although it does not seem to carry the MOPC 173 variable region gene. These observations suggest that somatic DNA rearrangement of immunoglobulin light chain genes can involve both homologous chromosomes.Images

MeSH Terms
Animals Cell Line DNA Restriction Enzymes DNA, Recombinant/metabolism Escherichia coli/metabolism Immunoglobulin Light Chains/biosynthesis Immunoglobulin kappa-Chains/biosynthesis Liver/metabolism Mice Nucleic Acid Hybridization Plasmacytoma Protein Biosynthesis Transcription, Genetic
Chemicals
DNA, Recombinant Immunoglobulin Light Chains Immunoglobulin kappa-Chains DNA Restriction Enzymes
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Steinmetz M
Zachau H G
Mach B
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33 references, click to expand
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Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
0305-1048
DOI
10.1093/nar/6.10.3213
Published
1979-07-25
Pages
3213-29
Language
English
Region
England
NLM ID
0411011
PMCID
PMC327930
Subset
IM
External Links
PubMed source DOI: 10.1093/nar/6.10.3213 PMC Full Text View journal details
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