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Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1978 Aug;75(8):3881–3885. doi: 10.1073/pnas.75.8.3881

Multiple related immunoglobulin variable-region genes identified by cloning and sequence analysis.

J G Seidman, A Leder, M H Edgell, F Polsky, S M Tilghman, D C Tiemeier, P Leder
PMCID: PMC392892  PMID: 279004

Abstract

We have identified at least six EcoRI fragments of mouse DNA that encode variable-region gene sequences closely related to the mouse kappa light chain, MOPC-149. Two of these fragments have been cloned, and the entire nucleotide sequence of the variable-region genes encoded on each has been determined. Both genes encode closely related variable-region sequences extending from codon position 1 through position 97. Neither fragment encodes a constant-region sequence. Although both genes are closely related, they differ from one another and from the sequence expressed in the MOPC-149 cell from which they were cloned. These few differences cluster within the complementarity-determining regions although several occur in framework sequences as well. We therefore conclude that an antibody-producing cell contains genetic information corresponding to its expressed sequence and several other closely related but silent sequences. These initial results raise the possibility that similar sets of genes might exist corresponding to each of the many subgroups already identified among mouse kappa light chains. If true, this would further suggest that the mouse genome might be rich enough in variable-region genes so as to encode a major portion of the variable-region repertoire.

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Selected References

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  1. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  2. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  3. Brack C., Tonegawa S. Variable and constant parts of the immunoglobulin light chain gene of a mouse myeloma cell are 1250 nontranslated bases apart. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5652–5656. doi: 10.1073/pnas.74.12.5652. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brenner S., Milstein C. Origin of antibody variation. Nature. 1966 Jul 16;211(5046):242–243. doi: 10.1038/211242a0. [DOI] [PubMed] [Google Scholar]
  5. Cohn M. The take-home lesson--1971. Ann N Y Acad Sci. 1971 Dec 31;190:529–584. doi: 10.1111/j.1749-6632.1971.tb13562.x. [DOI] [PubMed] [Google Scholar]
  6. Denhardt D. T. A membrane-filter technique for the detection of complementary DNA. Biochem Biophys Res Commun. 1966 Jun 13;23(5):641–646. doi: 10.1016/0006-291x(66)90447-5. [DOI] [PubMed] [Google Scholar]
  7. Dreyer W. J., Bennett J. C. The molecular basis of antibody formation: a paradox. Proc Natl Acad Sci U S A. 1965 Sep;54(3):864–869. doi: 10.1073/pnas.54.3.864. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Edelman G. M., Gally J. A. Somatic recombination of duplicated genes: an hypothesis on the origin of antibody diversity. Proc Natl Acad Sci U S A. 1967 Feb;57(2):353–358. doi: 10.1073/pnas.57.2.353. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Farace M. G., Aellen M. F., Briand P. A., Faust C. H., Vassalli P., Mach B. No detectable reiteration of genes coding for mouse MOPC 41 immunoglobulin light-chain mRNA. Proc Natl Acad Sci U S A. 1976 Mar;73(3):727–731. doi: 10.1073/pnas.73.3.727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Faust C. H., Diggelmann H., Mach B. Estimation of the number of genes coding for the constant part of the mouse immunoglobulin kappa light chain. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2491–2495. doi: 10.1073/pnas.71.6.2491. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gally J. A., Edelman G. M. Somatic translocation of antibody genes. Nature. 1970 Jul 25;227(5256):341–348. doi: 10.1038/227341a0. [DOI] [PubMed] [Google Scholar]
  12. Honjo T., Packman S. Quantitation of constant and variable region genes for mouse immunoglobulin lambda chains. Biochemistry. 1976 Jun 29;15(13):2780–2785. doi: 10.1021/bi00658a012. [DOI] [PubMed] [Google Scholar]
  13. Honjo T., Packman S., Swan D., Nau M., Leder P. Organization of immunoglobulin genes: reiteration frequency of the mouse kappa chain constant region gene. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3659–3663. doi: 10.1073/pnas.71.9.3659. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Honjo T., Swan D., Packman S., Polsky F., Leder P. Purification and translation of an immunoglobulin lambda chain messenger RNA from mouse myeloma. Biochemistry. 1976 Jun 29;15(13):2775–2779. doi: 10.1021/bi00658a011. [DOI] [PubMed] [Google Scholar]
  15. Hood L., McKean D., Farnsworth V., Potter M. Mouse immunoglobulin chains. A survey of the amino-terminal sequences of kappa chains. Biochemistry. 1973 Feb;12(4):741–749. doi: 10.1021/bi00728a026. [DOI] [PubMed] [Google Scholar]
  16. Hozumi N., Tonegawa S. Evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3628–3632. doi: 10.1073/pnas.73.10.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jerne N. K. The somatic generation of immune recognition. Eur J Immunol. 1971 Jan;1(1):1–9. doi: 10.1002/eji.1830010102. [DOI] [PubMed] [Google Scholar]
  18. LEDERBERG J. Genes and antibodies. Science. 1959 Jun 19;129(3364):1649–1653. doi: 10.1126/science.129.3364.1649. [DOI] [PubMed] [Google Scholar]
  19. Leder P., Honjo T., Packman S., Swan D., Nau M., Norman B. The organization and diversity of immunoglobulin genes. Proc Natl Acad Sci U S A. 1974 Dec;71(12):5109–5115. doi: 10.1073/pnas.71.12.5109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Milstein C., Brownlee G. G., Harrison T. M., Mathews M. B. A possible precursor of immunoglobulin light chains. Nat New Biol. 1972 Sep 27;239(91):117–120. doi: 10.1038/newbio239117a0. [DOI] [PubMed] [Google Scholar]
  22. Polsky F., Edgell M. H., Seidman J. G., Leder P. High capacity gel preparative electrophoresis for purification of fragments of genomic DNA. Anal Biochem. 1978 Jul 1;87(2):397–410. doi: 10.1016/0003-2697(78)90689-9. [DOI] [PubMed] [Google Scholar]
  23. Rabbitts T. H., Jarvis J. M., Milstein C. Demonstration that a mouse immunoglobulin light chain messenger RNA hybridizes exclusively with unique DNA. Cell. 1975 Sep;6(1):5–12. doi: 10.1016/0092-8674(75)90067-7. [DOI] [PubMed] [Google Scholar]
  24. Rabbitts T. H., Milstein C. Mouse immunoglobulin genes: studies on the reiteration frequency of light-chain genes by hybridisation procedures. Eur J Biochem. 1975 Mar 3;52(1):125–133. doi: 10.1111/j.1432-1033.1975.tb03980.x. [DOI] [PubMed] [Google Scholar]
  25. Rose S. M., Kuehl W. M., Smith G. P. Cloned MPC 11 myeloma cells express two kappa genes: a gene for a complete light chain and a gene for a constant region polypeptide. Cell. 1977 Oct;12(2):453–462. doi: 10.1016/0092-8674(77)90121-0. [DOI] [PubMed] [Google Scholar]
  26. SMITHIES O. GAMMA-GLOBULIN VARIABILITY: A GENETIC HYPOTHESIS. Nature. 1963 Sep 28;199:1231–1236. doi: 10.1038/1991231a0. [DOI] [PubMed] [Google Scholar]
  27. Seidman J. G., Edgell M. H., Leder P. Immunoglobulin light-chain structural gene sequences cloned in a bacterial plasmid. Nature. 1978 Feb 9;271(5645):582–585. doi: 10.1038/271582a0. [DOI] [PubMed] [Google Scholar]
  28. Smithies O. Antibody variability. Somatic recombination between the elements of "antibody gene pairs" may explain antibody variability. Science. 1967 Jul 21;157(3786):267–273. doi: 10.1126/science.157.3786.267. [DOI] [PubMed] [Google Scholar]
  29. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  30. Stavnezer J., Huang R. C., Stavnezer E., Bishop J. M. Isolation of messenger RNA for an immunoglobulin kappa chain and enumeration of the genes for the constatn region of kappa chain in the mouse. J Mol Biol. 1974 Sep 5;88(1):43–63. doi: 10.1016/0022-2836(74)90294-0. [DOI] [PubMed] [Google Scholar]
  31. Storb U. Evidence for multiple immunoglobulin genes. Biochem Biophys Res Commun. 1974 Mar 15;57(1):31–38. doi: 10.1016/s0006-291x(74)80353-0. [DOI] [PubMed] [Google Scholar]
  32. Swan D., Aviv H., Leder P. Purification and properties of biologically active messenger RNA for a myeloma light chain. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1967–1971. doi: 10.1073/pnas.69.7.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Szilard L. THE MOLECULAR BASIS OF ANTIBODY FORMATION. Proc Natl Acad Sci U S A. 1960 Mar;46(3):293–302. doi: 10.1073/pnas.46.3.293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tiemeier D. C., Tilghman S. M., Leder P. Purification and cloning of a mouse ribosomal gene fragment in coliphage lambda. Gene. 1977;2(3-4):173–191. doi: 10.1016/0378-1119(77)90016-6. [DOI] [PubMed] [Google Scholar]
  35. Tilghman S. M., Tiemeier D. C., Polsky F., Edgell M. H., Seidman J. G., Leder A., Enquist L. W., Norman B., Leder P. Cloning specific segments of the mammalian genome: bacteriophage lambda containing mouse globin and surrounding gene sequences. Proc Natl Acad Sci U S A. 1977 Oct;74(10):4406–4410. doi: 10.1073/pnas.74.10.4406. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Tonegawa S., Brack C., Hozumi N., Schuller R. Cloning of an immunoglobulin variable region gene from mouse embryo. Proc Natl Acad Sci U S A. 1977 Aug;74(8):3518–3522. doi: 10.1073/pnas.74.8.3518. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tonegawa S., Hozumi N., Matthyssens G., Schuller R. Somatic changes in the content and context of immunoglobulin genes. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):877–889. doi: 10.1101/sqb.1977.041.01.097. [DOI] [PubMed] [Google Scholar]
  38. Tonegawa S., Maxam A. M., Tizard R., Bernard O., Gilbert W. Sequence of a mouse germ-line gene for a variable region of an immunoglobulin light chain. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1485–1489. doi: 10.1073/pnas.75.3.1485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tonegawa S. Reiteration frequency of immunoglobulin light chain genes: further evidence for somatic generation of antibody diversity. Proc Natl Acad Sci U S A. 1976 Jan;73(1):203–207. doi: 10.1073/pnas.73.1.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Tonegawa S., Steinberg C., Dube S., Bernardini A. Evidence for somatic generation of antibody diversity. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4027–4031. doi: 10.1073/pnas.71.10.4027. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. White R. L., Hogness D. S. R loop mapping of the 18S and 28S sequences in the long and short repeating units of Drosophila melanogaster rDNA. Cell. 1977 Feb;10(2):177–192. doi: 10.1016/0092-8674(77)90213-6. [DOI] [PubMed] [Google Scholar]

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