Immunoglobulin heavy chain gene organization in mice: Analysis of a myeloma genomic clone containing variable and α constant regions

Creators: Early, Philip W.; Davis, Mark M.; Kaback, David B.; Davidson, Norman; Hood, Leroy

Abstract

We have isolated a myeloma genomic DNA clone containing the variable and constant regions of a mouse chain. Restriction enzyme analyses and electron microscopic R loop mapping have demonstrated that the variable region is separated from the constant region by 6.8 kilobases of intervening DNA. In addition, two intervening DNA sequences of 100-200 bases separate the constant region into three approximately equal units. These intervening sequences may separate each of the segments coding for the three constant region domains of the heavy chain. Southern blot analysis of embryo and myeloma DNA suggests that DNA rearrangement of heavy chain variable and constant regions occurs during the differentiation of antibody-producing cells.

Additional Information

© 1979 by the National Academy of Sciences. Contributed by Norman Davidson, November 7, 1978. We thank H. Manor for electron microscopy of p603α1, N. Johnson for providing M603 DNA, and Y.-H. Chien for electron microscopic characterization of mRNAs. E. coli polymerase I, DNA ligase T4, and EcoRI linkers were generous gifts from M. Goldberg, R. Scheller, and K. Itakura, respectively. Most of the in vitro packaging extracts used were the gift of T. Sargent. We thank K. Marcu, O. Valbuena, and R. Perry for advice on mRNA preparation and M. Wickens for communicating cDNA synthesis procedures prior to publication. We benefited from helpful discussions with T. Maniatis, T. Sargent, D. Goldberg, D. Engel, J. Dodgson, R. Joho, B. Klein, and D. Anderson. The work reported here was supported by National Institutes of Health Grants GM 10991 and GM 20927 and Biomedical Research Grant RRO7003A and National Science Foundation Grant PCM 71-00770. P.W.E. and M.M.D. are supported by National Institutes of Health Training Grant GM 07616; D.B.K. is a National Institutes of Health Postdoctoral Fellow. The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.

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