Skip to main content
PMC home page
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
. 1980 Oct;77(10):6027–6031. doi: 10.1073/pnas.77.10.6027

Cloning and partial nucleotide sequence of human immunoglobulin mu chain cDNA from B cells and mouse-human hybridomas.

T W Dolby, J Devuono, C M Croce
PMCID: PMC350206  PMID: 6777778

Abstract

Purified mRNAs coding for mu and kappa human immunoglobulin polypeptides were translated in vitro and their products were characterized. The mu-specific mRNAs, derived from both human lymphoblastoid cells (GM607) and from a mouse-human somatic cell hybrid secreting human mu chains (alpha D5-H11-BC11), were copied into cDNAs and inserted into the plasmid pBR322. Several recombinant cDNAs that were obtained were identified by a combination of colony hybridization with labeled probes, in vitro translation of plasmid-selected mu mRNAs, and DNA nucleotide sequence determination. One recombinant DNA, for which the sequence has been partially determined, contains the codons for part of the C3 constant region domain through the carboxy-terminal piece (155 amino acids total) as well as the entire 3' noncoding sequence up to the poly(A) site of the human mu mRNA. The sequence A-A-U-A-A occurs 12 nucleotides prior to the poly(A) addition site in the human mu mRNA. Considerable sequence homology is observed in the mouse and human mu mRNA 3' coding and noncoding sequences.

Full text

PDF
6031

Images in this article

6028Image
on p.6028
6028Image
on p.6028
6029Image
on p.6029

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burstein Y., Schechter I. Amino acid sequence of the NH2-terminal extra piece segments of the precursors of mouse immunoglobulin lambda1-type and kappa-type light chains. Proc Natl Acad Sci U S A. 1977 Feb;74(2):716–720. doi: 10.1073/pnas.74.2.716. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Calame K., Rogers J., Early P., Davis M., Livant D., Wall R., Hood L. Mouse Cmu heavy chain immunoglobulin gene segment contains three intervening sequences separating domains. Nature. 1980 Apr 3;284(5755):452–455. doi: 10.1038/284452a0. [DOI] [PubMed] [Google Scholar]
  3. Croce C. M., Shander M., Martinis J., Cicurel L., D'Ancona G. G., Dolby T. W., Koprowski H. Chromosomal location of the genes for human immunoglobulin heavy chains. Proc Natl Acad Sci U S A. 1979 Jul;76(7):3416–3419. doi: 10.1073/pnas.76.7.3416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. 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]
  5. Enea V., Vovis G. F., Zinder N. D. Genetic studies with heteroduplex DNA of bacteriophage fl. Asymmetric segregation, base correction and implications for the mechanism of genetic recombination. J Mol Biol. 1975 Aug 15;96(3):495–509. doi: 10.1016/0022-2836(75)90175-8. [DOI] [PubMed] [Google Scholar]
  6. Goldberg M. L., Lifton R. P., Stark G. R., Williams J. G. Isolation of specific RNA's using DNA covalently linked to diazobenzyloxymethyl cellulose or paper. Methods Enzymol. 1979;68:206–220. doi: 10.1016/0076-6879(79)68016-3. [DOI] [PubMed] [Google Scholar]
  7. Honjo T., Obata M., Yamawaki-Katoaka Y., Kataoka T., Kawakami T., Takahashi N., Mano Y. Cloning and complete nucleotide sequence of mouse immunoglobulin gamma 1 chain gene. Cell. 1979 Oct;18(2):559–568. doi: 10.1016/0092-8674(79)90072-2. [DOI] [PubMed] [Google Scholar]
  8. Horibata K., Harris A. W. Mouse myelomas and lymphomas in culture. Exp Cell Res. 1970 Apr;60(1):61–77. doi: 10.1016/0014-4827(70)90489-1. [DOI] [PubMed] [Google Scholar]
  9. Humphries P., Old R., Coggins L. W., McShane T., Watson C., Paul J. Recombinant plasmids containing Xenopus laevis globin structural genes derived from complementary DNA. Nucleic Acids Res. 1978 Mar;5(3):905–924. doi: 10.1093/nar/5.3.905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jilka R. L., Pestka S. Amino acid sequence of the precursor region of MOPC-315 mouse immunoglobulin heavy chain. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5692–5696. doi: 10.1073/pnas.74.12.5692. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kafatos F. C., Efstratiadis A., Forget B. G., Weissman S. M. Molecular evolution of human and rabbit beta-globin mRNAs. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5618–5622. doi: 10.1073/pnas.74.12.5618. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  13. Klukas C. K., Cramer F., Gould H. Translation of human immunoglobulin heavy chain mRNA in vitro. Nature. 1977 Sep 15;269(5625):262–264. doi: 10.1038/269262a0. [DOI] [PubMed] [Google Scholar]
  14. Konkel D. A., Tilghman S. M., Leder P. The sequence of the chromosomal mouse beta-globin major gene: homologies in capping, splicing and poly(A) sites. Cell. 1978 Dec;15(4):1125–1132. doi: 10.1016/0092-8674(78)90040-5. [DOI] [PubMed] [Google Scholar]
  15. Kupersztoch Y. M., Helinski D. R. A catenated DNA molecule as an intermediate in the replication of the resistance transfer factor R6K in Escherichia coli. Biochem Biophys Res Commun. 1973 Oct 15;54(4):1451–1459. doi: 10.1016/0006-291x(73)91149-2. [DOI] [PubMed] [Google Scholar]
  16. Maniatis T., Jeffrey A., van deSande H. Chain length determination of small double- and single-stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry. 1975 Aug 26;14(17):3787–3794. doi: 10.1021/bi00688a010. [DOI] [PubMed] [Google Scholar]
  17. Marcu K. B., Valbuena O., Perry R. P. Isolation, purification, and properties of mouse heavy-chain immunoglobulin mRNAs. Biochemistry. 1978 May 2;17(9):1723–1733. doi: 10.1021/bi00602a022. [DOI] [PubMed] [Google Scholar]
  18. Matthyssens G., Rabbitts T. H. The sequence at the 3' terminus of mouse immunoglobulin secreted mu chain messenger RNA determined from cloned cDNA. Nucleic Acids Res. 1980 Feb 25;8(4):703–713. [PMC free article] [PubMed] [Google Scholar]
  19. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  20. Myers J. C., Spiegelman S., Kacian D. L. Synthesis of full-length DNA copies of avian myeloblastosis virus RNA in high yields. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2840–2843. doi: 10.1073/pnas.74.7.2840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pelham H. R., Jackson R. J. An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem. 1976 Aug 1;67(1):247–256. doi: 10.1111/j.1432-1033.1976.tb10656.x. [DOI] [PubMed] [Google Scholar]
  22. Potter M. Immunoglobulin-producing tumors and myeloma proteins of mice. Physiol Rev. 1972 Jul;52(3):631–719. doi: 10.1152/physrev.1972.52.3.631. [DOI] [PubMed] [Google Scholar]
  23. Putnam F. W., Florent G., Paul C., Shinoda T., Shimizu A. Complete amino acid sequence of the Mu heavy chain of a human IgM immunoglobulin. Science. 1973 Oct 19;182(4109):287–291. doi: 10.1126/science.182.4109.287. [DOI] [PubMed] [Google Scholar]
  24. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  25. Roychoudhury R., Jay E., Wu R. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res. 1976 Jan;3(1):101–116. doi: 10.1093/nar/3.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Seeburg P. H., Shine J., Martial J. A., Baxter J. D., Goodman H. M. Nucleotide sequence and amplification in bacteria of structural gene for rat growth hormone. Nature. 1977 Dec 8;270(5637):486–494. doi: 10.1038/270486a0. [DOI] [PubMed] [Google Scholar]
  27. Shenk T. E., Rhodes C., Rigby P. W., Berg P. Biochemical method for mapping mutational alterations in DNA with S1 nuclease: the location of deletions and temperature-sensitive mutations in simian virus 40. Proc Natl Acad Sci U S A. 1975 Mar;72(3):989–993. doi: 10.1073/pnas.72.3.989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Sippel A. E., Land H., Lindenmaier W., Nguyen-Huu M. C., Wurtz T., Timmis K. N., Giesecke K., Schütz G. Cloning of chicken lysozyme structural gene sequences synthesized in vitro. Nucleic Acids Res. 1978 Sep;5(9):3275–3294. doi: 10.1093/nar/5.9.3275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Stephens R. E., Pan C. J., Ajiro K., Dolby T. W., Borun T. W. Studies of human histone messenger RNA. I. Methods for the isolation and partial characterization of RNA fractions containing human histone message from HeLa S3 polyribosomes. J Biol Chem. 1977 Jan 10;252(1):166–172. [PubMed] [Google Scholar]
  30. Villa-Komaroff L., Efstratiadis A., Broome S., Lomedico P., Tizard R., Naber S. P., Chick W. L., Gilbert W. A bacterial clone synthesizing proinsulin. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3727–3731. doi: 10.1073/pnas.75.8.3727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Watanabe S., Barnikol H. U., Horn J., Bertram J., Hilschmann N. Die Primärstruktur eines monoklonalen IgM-Immunoglobulins (Makroglobulin Gal.), II. Die Aminosäuresequenz der H-Kette (mu-Typ,Subgruppe HIII), Struktur des gesamten IgM-Moleküls. Hoppe Seylers Z Physiol Chem. 1973 Oct-Nov;354(10-11):1505–1509. [PubMed] [Google Scholar]
  32. Wickens M. P., Buell G. N., Schimke R. T. Synthesis of double-stranded DNA complementary to lysozyme, ovomucoid, and ovalbumin mRNAs. Optimization for full length second strand synthesis by Escherichia coli DNA polymerase I. J Biol Chem. 1978 Apr 10;253(7):2483–2495. [PubMed] [Google Scholar]
  33. Williams P. B., Kubo R. T., Grey H. M. mu-Chains from a nonsecretor B cell line differ from secreted mu-chains at the C-terminal end. J Immunol. 1978 Dec;121(6):2435–2439. [PubMed] [Google Scholar]
  34. Yaffe L., Pestka S. Isolation and cell-free translation of a human immunoglobulin light chain messenger RNA. Arch Biochem Biophys. 1978 Oct;190(2):495–507. doi: 10.1016/0003-9861(78)90303-x. [DOI] [PubMed] [Google Scholar]
  35. Zakut R., Givol D., Mory Y. Y. Structure of immunoglobulin gamma 2b heavy chain gene cloned from mouse embryo gene library. Nucleic Acids Res. 1980 Feb 11;8(3):453–466. doi: 10.1093/nar/8.3.453. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES