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. 1982 Mar 11;10(5):1495–1502. doi: 10.1093/nar/10.5.1495

Chicken histone H5: selection of a cDNA recombinant using an extended synthetic primer.

P A Krieg, A J Robins, M J Gait, R C Titmas, J R Wells
PMCID: PMC320545  PMID: 6896089

Abstract

We describe the use of a synthetic primer to select a cDNA recombinant clone containing H5 coding sequences. The strategy used was as follows: 1. Prepare oligo(dT) cellulose-bound mRNA from chicken reticulocytes and select 11S-18S material from sucrose gradients. 2. Use this RNA fraction both to prepare a cDNA library and as a template for H5-specific cDNA synthesis using a synthetic primer. 3. Screen out most globin cDNA recombinants with oligo(dT)-primed globin cDNA. 4. Search for H5 recombinants using H5 specific cDNA and verify the identity by DNA sequencing. Our screening suggests an H5 mRNA abundance of about two parts per thousand in chicken reticulocyte poly(A)-containing RNA. The isolation of an H5 cDNA recombinant clone is an initial step in the study of H5 genes and their relationship to H1 and core histone genes.

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Selected References

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  1. Appels R., Wells J. R. Synthesis and turnover of DNA-bound histone during maturation of avian red blood cells. J Mol Biol. 1972 Oct 14;70(3):425–434. doi: 10.1016/0022-2836(72)90550-5. [DOI] [PubMed] [Google Scholar]
  2. Appels R., Wells J. R., Williams A. F. Characterization of DNA-bound histone in the cells of the avian erythropoietic series. J Cell Sci. 1972 Jan;10(1):47–59. doi: 10.1242/jcs.10.1.47. [DOI] [PubMed] [Google Scholar]
  3. Billett M. A., Hindley J. A study of the quantitative variation of histones, and their relationship to RNA synthesis, during erythropoiesis in the adult chicken. Eur J Biochem. 1972 Aug 4;28(4):451–462. doi: 10.1111/j.1432-1033.1972.tb01932.x. [DOI] [PubMed] [Google Scholar]
  4. Briand G., Kmiecik D., Sautiere P., Wouters D., Borie-Loy O., Biserte G., Mazen A., Champagne M. Chicken erythrocyte histone H5. IV. Sequence of the carboxy-termined half of the molecule (96 residues) and complete sequence. FEBS Lett. 1980 Apr 7;112(2):147–151. doi: 10.1016/0014-5793(80)80167-0. [DOI] [PubMed] [Google Scholar]
  5. Chang A. C., Nunberg J. H., Kaufman R. J., Erlich H. A., Schimke R. T., Cohen S. N. Phenotypic expression in E. coli of a DNA sequence coding for mouse dihydrofolate reductase. Nature. 1978 Oct 19;275(5681):617–624. doi: 10.1038/275617a0. [DOI] [PubMed] [Google Scholar]
  6. Duckworth M. L., Gait M. J., Goelet P., Hong G. F., Singh M., Titmas R. C. Rapid synthesis of oligodeoxyribonucleotides VI. Efficient, mechanised synthesis of heptadecadeoxyribonucleotides by an improved solid phase phosphotriester route. Nucleic Acids Res. 1981 Apr 10;9(7):1691–1706. doi: 10.1093/nar/9.7.1691. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grunstein M., Wallis J. Colony hybridization. Methods Enzymol. 1979;68:379–389. doi: 10.1016/0076-6879(79)68027-8. [DOI] [PubMed] [Google Scholar]
  8. Houghton M., Stewart A. G., Doel S. M., Emtage J. S., Eaton M. A., Smith J. C., Patel T. P., Lewis H. M., Porter A. G., Birch J. R. The amino-terminal sequence of human fibroblast interferon as deduced from reverse transcripts obtained using synthetic oligonucleotide primers. Nucleic Acids Res. 1980 May 10;8(9):1913–1931. doi: 10.1093/nar/8.9.1913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hudson P., Haley J., Cronk M., Shine J., Niall H. Molecular cloning and characterization of cDNA sequences coding for rat relaxin. Nature. 1981 May 14;291(5811):127–131. doi: 10.1038/291127a0. [DOI] [PubMed] [Google Scholar]
  10. Kornberg R. The location of nucleosomes in chromatin: specific or statistical. Nature. 1981 Aug 13;292(5824):579–580. doi: 10.1038/292579a0. [DOI] [PubMed] [Google Scholar]
  11. Molgaard H. V., Perucho M., Ruiz-Carrillo A. Histone H5 messenger RNA is polyadenylated. Nature. 1980 Jan 31;283(5746):502–504. doi: 10.1038/283502a0. [DOI] [PubMed] [Google Scholar]
  12. Newrock K. M., Alfageme C. R., Nardi R. V., Cohen L. H. Histone changes during chromatin remodeling in embryogenesis. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 1):421–431. doi: 10.1101/sqb.1978.042.01.045. [DOI] [PubMed] [Google Scholar]
  13. Richards R. I., Shine J., Ullrich A., Wells J. R., Goodman H. M. Molecular cloning and sequence analysis of adult chicken betal globin cDNA. Nucleic Acids Res. 1979 Nov 10;7(5):1137–1146. doi: 10.1093/nar/7.5.1137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Richards R. I., Wells J. R. Chicken globin genes. Nucleotide sequence of cDNA clones coding for the alpha-globin expressed during hemolytic anemia. J Biol Chem. 1980 Oct 10;255(19):9306–9311. [PubMed] [Google Scholar]
  15. 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 Apr;3(4):863–877. doi: 10.1093/nar/3.4.863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Ruiz-Carrillo A., Wangh L. J., Littau V. C., Allfrey V. G. Changes in histone acetyl content and in nuclear non-histone protein composition of avian erythroid cells at different stages of maturation. J Biol Chem. 1974 Nov 25;249(22):7358–7368. [PubMed] [Google Scholar]
  17. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Savić A., Richman P., Williamson P., Poccia D. Alterations in chromatin structure during early sea urchin embryogenesis. Proc Natl Acad Sci U S A. 1981 Jun;78(6):3706–3710. doi: 10.1073/pnas.78.6.3706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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