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. 1978 Nov;75(11):5367–5371. doi: 10.1073/pnas.75.11.5367

Molecular analysis of cloned human 18S ribosomal DNA segments.

G N Wilson, B A Hollar, J R Waterson, R D Schmickel
PMCID: PMC392964  PMID: 364477

Abstract

A fraction of DNA from the human fetal lung fibroblast line IMR-90, 30-fold enriched for ribosomal DNA, was cloned in the lambda phage vector Charon 16A. Of 978 clones assayed by hybridization to a mixture of 125I-labeled 18S and 28S ribosomal RNA, 11 recombinants containing a 3.8-megadalton segment of human 18S ribosomal DNA were identified. Restriction endonuclease analysis of these clones demonstrated variation only in orientation of the human gene segment within the phage vector. Restriction sites that we had previously detected from analysis of restriction products of unfractionated human DNA by using the Southern transfer method were also present in the cloned DNA segment. Recombinant DNA technology thus provides a valid and efficient means to define structural conservation or variation within families of human genes.

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Selected References

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  1. Arnheim N., Southern E. M. Heterogeneity of the ribosomal genes in mice and men. Cell. 1977 Jun;11(2):363–370. doi: 10.1016/0092-8674(77)90053-8. [DOI] [PubMed] [Google Scholar]
  2. Blattner F. R., Williams B. G., Blechl A. E., Denniston-Thompson K., Faber H. E., Furlong L., Grunwald D. J., Kiefer D. O., Moore D. D., Schumm J. W. Charon phages: safer derivatives of bacteriophage lambda for DNA cloning. Science. 1977 Apr 8;196(4286):161–169. doi: 10.1126/science.847462. [DOI] [PubMed] [Google Scholar]
  3. Bross K., Krone W. Ribosomal cistrons and acrocentric chromosomes in man. Humangenetik. 1973 Mar 23;18(1):71–75. doi: 10.1007/BF00279033. [DOI] [PubMed] [Google Scholar]
  4. Helling R. B., Goodman H. M., Boyer H. W. Analysis of endonuclease R-EcoRI fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J Virol. 1974 Nov;14(5):1235–1244. doi: 10.1128/jvi.14.5.1235-1244.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. LENNOX E. S. Transduction of linked genetic characters of the host by bacteriophage P1. Virology. 1955 Jul;1(2):190–206. doi: 10.1016/0042-6822(55)90016-7. [DOI] [PubMed] [Google Scholar]
  6. Laskey R. A., Mills A. D. Quantitative film detection of 3H and 14C in polyacrylamide gels by fluorography. Eur J Biochem. 1975 Aug 15;56(2):335–341. doi: 10.1111/j.1432-1033.1975.tb02238.x. [DOI] [PubMed] [Google Scholar]
  7. Mandel M., Higa A. Calcium-dependent bacteriophage DNA infection. J Mol Biol. 1970 Oct 14;53(1):159–162. doi: 10.1016/0022-2836(70)90051-3. [DOI] [PubMed] [Google Scholar]
  8. Miller O. L., Jr, Bakken A. H. Morphological studies of transcription. Acta Endocrinol Suppl (Copenh) 1972;168:155–177. doi: 10.1530/acta.0.071s155. [DOI] [PubMed] [Google Scholar]
  9. Murray K. Nucleotide sequence analysis with polynucleotide kinase and nucleotide "mapping" methods. 5'-Terminal sequences of deoxyribonucleic acid from bacteriophages lambda and 424. Biochem J. 1973 Mar;131(3):569–582. doi: 10.1042/bj1310569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Nichols W. W., Murphy D. G., Cristofalo V. J., Toji L. H., Greene A. E., Dwight S. A. Characterization of a new human diploid cell strain, IMR-90. Science. 1977 Apr 1;196(4285):60–63. doi: 10.1126/science.841339. [DOI] [PubMed] [Google Scholar]
  11. Schmickel R. D., Knoller M. Characterization and localization of the human genes for ribosomal ribonucleic acid. Pediatr Res. 1977 Aug;11(8):929–935. doi: 10.1203/00006450-197708000-00015. [DOI] [PubMed] [Google Scholar]
  12. Schmickel R. D. Quantitation of human ribosomal DNA: hybridization of human DNA with ribosomal RNA for quantitation and fractionation. Pediatr Res. 1973 Jan;7(1):5–12. doi: 10.1203/00006450-197301000-00002. [DOI] [PubMed] [Google Scholar]
  13. 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]
  14. Young B. D., Hell A., Birnie G. D. A new estimate of human ribosomal gene number. Biochim Biophys Acta. 1976 Dec 13;454(3):539–548. doi: 10.1016/0005-2787(76)90279-3. [DOI] [PubMed] [Google Scholar]

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