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. 1979 Apr;6(4):1241–1258. doi: 10.1093/nar/6.4.1241

A precise quantitation of gene number by saturation hybridization using cloned DNA.

S S Longacre, B Mach
PMCID: PMC327770  PMID: 450695

Abstract

This paper describes a precise method of gene titration as applied to the alpha- and beta-globin genes in the mouse. The three salient features of the method are: (i) the use of saturation hybridization in probe cDNA excess, (ii) the use of highly purified cDNA probes prepared by preparative hybridization with cloned globin sequences (Longacre and Mach (1978) J. Biol. Chem. 253, 7500) and (iii) the use of cloned globin sequences to calibrate the system internally. The results indicate that there are two genes for alpha-globin and two genes for beta-globin in the BALB/c mouse. The significance of these results are discussed in relation to other data regarding adult and embryonic globin genes.

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Selected References

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  1. Bishop J. O., Freeman K. B. DNA sequences neighboring the duck hemoglobin genes. Cold Spring Harb Symp Quant Biol. 1974;38:707–716. doi: 10.1101/sqb.1974.038.01.076. [DOI] [PubMed] [Google Scholar]
  2. Bonner T. I., Brenner D. J., Neufeld B. R., Britten R. J. Reduction in the rate of DNA reassociation by sequence divergence. J Mol Biol. 1973 Dec 5;81(2):123–135. doi: 10.1016/0022-2836(73)90184-8. [DOI] [PubMed] [Google Scholar]
  3. Britten R. J., Kohne D. E. Repeated sequences in DNA. Hundreds of thousands of copies of DNA sequences have been incorporated into the genomes of higher organisms. Science. 1968 Aug 9;161(3841):529–540. doi: 10.1126/science.161.3841.529. [DOI] [PubMed] [Google Scholar]
  4. Chambon P. Summary: the molecular biology of the eukaryotic genome is coming of age. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):1209–1234. doi: 10.1101/sqb.1978.042.01.122. [DOI] [PubMed] [Google Scholar]
  5. Clegg J. B., Naughton M. A., Weatherball D. J. Abnormal human haemoglobins. Separation and characterization of the alpha and beta chains by chromatography, and the determination of two new variants, hb Chesapeak and hb J (Bangkok). J Mol Biol. 1966 Aug;19(1):91–108. doi: 10.1016/s0022-2836(66)80052-9. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Hilse K., Popp R. A. Gene duplication as the basis for amino acid ambiguity in the alpha-chain polypeptides of mouse hemoglobins. Proc Natl Acad Sci U S A. 1968 Nov;61(3):930–936. doi: 10.1073/pnas.61.3.930. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Katz L., Kingsbury D. T., Helinski D. R. Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid-protein relaxation complex. J Bacteriol. 1973 May;114(2):577–591. doi: 10.1128/jb.114.2.577-591.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Longacre S. S., Mach B. Purification of specific DNA sequences by sulfhydryl-Sepharose chromatography of mercurated polynucleotides. J Biol Chem. 1978 Oct 25;253(20):7500–7507. [PubMed] [Google Scholar]
  11. Melderis H., Steinheider G., Ostertag W. Evidence for a unique kind of alpha-type globin chain in early mammalian embryos. Nature. 1974 Aug 30;250(5469):774–776. doi: 10.1038/250774a0. [DOI] [PubMed] [Google Scholar]
  12. Popp R. A., Bailiff E. G. Sequence of amino acids in the major and minor chains of the diffuse hemoglobin from BALB-c mice. Biochim Biophys Acta. 1973 Mar 23;303(1):61–67. doi: 10.1016/0005-2795(73)90148-7. [DOI] [PubMed] [Google Scholar]
  13. Popp R. A. Hemoglobins of mice: sequence and possible ambiguity at one position of the alpha chain. J Mol Biol. 1967 Jul 14;27(1):9–16. doi: 10.1016/0022-2836(67)90347-6. [DOI] [PubMed] [Google Scholar]
  14. Popp R. A. Sequence of amino acids in the chain of single hemoglobins from C57BL, SWR, and NB mice. Biochim Biophys Acta. 1973 Mar 23;303(1):52–60. doi: 10.1016/0005-2795(73)90147-5. [DOI] [PubMed] [Google Scholar]
  15. Rougeon F., Kourilsky P., Mach B. Insertion of a rabbit beta-globin gene sequence into an E. coli plasmid. Nucleic Acids Res. 1975 Dec;2(12):2365–2378. doi: 10.1093/nar/2.12.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Rougeon F., Mach B. Cloning and amplification of alpha and beta mouse globin gene sequences synthesised in vitro. Gene. 1977 May;1(3-4):229–239. doi: 10.1016/0378-1119(77)90047-6. [DOI] [PubMed] [Google Scholar]
  17. 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]
  18. Steinheider G., Melderis H., Ostertag W. Embryonic epsilon chains of mice and rabbits. Nature. 1975 Oct 23;257(5528):714–716. doi: 10.1038/257714a0. [DOI] [PubMed] [Google Scholar]
  19. Steinheider G., Melderis H., Ostertag W. Evidence for Hb Lepore-like hybrid globin beta chain genes in mice. Nature. 1975 Oct 23;257(5528):712–714. doi: 10.1038/257712a0. [DOI] [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. 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]
  23. Vogt V. M. Purification and further properties of single-strand-specific nuclease from Aspergillus oryzae. Eur J Biochem. 1973 Feb 15;33(1):192–200. doi: 10.1111/j.1432-1033.1973.tb02669.x. [DOI] [PubMed] [Google Scholar]

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