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. 1978 Mar;75(3):1299–1303. doi: 10.1073/pnas.75.3.1299

Intragenic DNA spacers interrupt the ovalbumin gene

Ruth Weinstock *, Raymond Sweet , Michael Weiss , Howard Cedar §, Richard Axel
PMCID: PMC411458  PMID: 274719

Abstract

We have performed restriction endonuclease mapping to examine the linear organization of the ovalbumin gene in chromosomal DNA. Treatment of genomic DNA with restriction endonucleases that do not cleave the ovalbumin mRNA sequence results in the generation of multiple DNA fragments capable of annealing with ovalbumin-specific probes in molecular hybridization reactions. These data strongly suggest that the linear order of DNA sequences coding for ovalbumin is interrupted by at least two intragenic DNA spacers absent from the corresponding RNA. At least one of these spacer sequences interrupts the coding sequence; therefore the chromosomal ovalbumin gene is not colinear with its translational product. We can discern no difference in the sequence organization about this gene in producer and nonproducer somatic cells, suggesting that the presence of intragenic spacers does not reflect that transcriptional activity of this gene. Furthermore, profiles obtained for gametes are identical to those observed for somatic cells, indicating that significant translocation during development is not responsible for the generation of this split sequence. Divergence of the intragenic spacer, however, is observed between individual chickens; thus multiple alleles may exist for this gene that are identifiable by differences in the organization of spacer with no apparent phenotypic evidence for their presence.

Keywords: restriction endonuclease mapping, intragenic spacers, gene organization

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Selected References

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  1. Axel R., Cedar H., Felsenfeld G. Synthesis of globin ribonucleic acid from duck-reticulocyte chromatin in vitro. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2029–2032. doi: 10.1073/pnas.70.7.2029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Breathnach R., Mandel J. L., Chambon P. Ovalbumin gene is split in chicken DNA. Nature. 1977 Nov 24;270(5635):314–319. doi: 10.1038/270314a0. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Doel M. T., Houghton M., Cook E. A., Carey N. H. The presence of ovalbumin mRNA coding sequences in multiple restriction fragments of chicken DNA. Nucleic Acids Res. 1977 Nov;4(11):3701–3713. doi: 10.1093/nar/4.11.3701. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Garel A., Axel R. Selective digestion of transcriptionally active ovalbumin genes from oviduct nuclei. Proc Natl Acad Sci U S A. 1976 Nov;73(11):3966–3970. doi: 10.1073/pnas.73.11.3966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Goodman H. M., Olson M. V., Hall B. D. Nucleotide sequence of a mutant eukaryotic gene: the yeast tyrosine-inserting ochre suppressor SUP4-o. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5453–5457. doi: 10.1073/pnas.74.12.5453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  8. Harris S. E., Means A. R., Mitchell W. M., O'Malley B. W. Synthesis of (3H)DNA complementary to ovalbumin messenger RNA: evidence for limited copies of the ovalbumin gene in chick oviduct. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3776–3780. doi: 10.1073/pnas.70.12.3776. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Humphries P., Cochet M., Krust A., Gerlinger P., Kourilsky P., Chambon P. Molecular cloning of extensive sequences of the in vitro synthesized chicken ovalbumin structural gene. Nucleic Acids Res. 1977 Jul;4(7):2389–2406. doi: 10.1093/nar/4.7.2389. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jeffreys A. J., Flavell R. A. The rabbit beta-globin gene contains a large large insert in the coding sequence. Cell. 1977 Dec;12(4):1097–1108. doi: 10.1016/0092-8674(77)90172-6. [DOI] [PubMed] [Google Scholar]
  11. Ketner G., Kelly T. J., Jr Integrated simian virus 40 sequences in transformed cell DNA: analysis using restriction endonucleases. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1102–1106. doi: 10.1073/pnas.73.4.1102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Maniatis T., Jeffrey A., Kleid D. G. Nucleotide sequence of the rightward operator of phage lambda. Proc Natl Acad Sci U S A. 1975 Mar;72(3):1184–1188. doi: 10.1073/pnas.72.3.1184. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Marshall A. J., Burgoyne L. A. Interpretation of the properties of chromatin extracts from mammalian nuclei. Nucleic Acids Res. 1976 Apr;3(4):1101–1110. doi: 10.1093/nar/3.4.1101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Monahan J. J., Woo S. L., Liarakos C. D., O'Malley B. W. Ovalbumin gene. Action of restriction endonucleases upon DNA coding sequence. J Biol Chem. 1977 Jul 10;252(13):4722–4728. [PubMed] [Google Scholar]
  15. 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]
  16. Sullivan D., Palacios R., Stavnezer J., Taylor J. M., Faras A. J., Kiely M. L., Summers N. M., Bishop J. M., Schimke R. T. Synthesis of a deoxyribonucleic acid sequence complementary to ovalbumin messenger ribonucleic acid and quantification of ovalbumin genes. J Biol Chem. 1973 Nov 10;248(21):7530–7539. [PubMed] [Google Scholar]
  17. Tilghman S. M., Curtis P. J., Tiemeier D. C., Leder P., Weissmann C. The intervening sequence of a mouse beta-globin gene is transcribed within the 15S beta-globin mRNA precursor. Proc Natl Acad Sci U S A. 1978 Mar;75(3):1309–1313. doi: 10.1073/pnas.75.3.1309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tilghman S. M., Tiemeier D. C., Seidman J. G., Peterlin B. M., Sullivan M., Maizel J. V., Leder P. Intervening sequence of DNA identified in the structural portion of a mouse beta-globin gene. Proc Natl Acad Sci U S A. 1978 Feb;75(2):725–729. doi: 10.1073/pnas.75.2.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Wigler M., Silverstein S., Lee L. S., Pellicer A., Cheng Y. c., Axel R. Transfer of purified herpes virus thymidine kinase gene to cultured mouse cells. Cell. 1977 May;11(1):223–232. doi: 10.1016/0092-8674(77)90333-6. [DOI] [PubMed] [Google Scholar]

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