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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
. 1979 Feb;76(2):695–699. doi: 10.1073/pnas.76.2.695

alpha-Fetoprotein and albumin genes of rats: no evidence for amplification-deletion or rearrangement in rat liver carcinogenesis.

J M Sala-Trepat, T D Sargent, S Sell, J Bonner
PMCID: PMC383021  PMID: 85303

Abstract

Full-length radiolabeled albumin and alpha-fetoprotein (AFP) cDNAs were synthesized from pure albumin and AMP mRNA preparations by using avian myeloblastosis virus reverse transcriptase (RNA-dependent DNA polymerase). The cDNAs have been used to quantitate the number of albumin and AFP genes in different rat tissues by two independent methods, both of which yielded similar results. First, the kinetics of the association of these cDNAs with nuclear DNA from rat liver, rat kidney, and Morris hepatoma 7777 under conditions of vast DNA excess indicated that the albumin and AFP mRNA's are transcribed from "nonrepetitive DNA." Second, saturation hybridization experiments in which a constant amount of rat liver DNA or Morris hepatoma 7777 was hybridized with increasing amounts of cDNA to albumin mRNA have shown the presence of 1--2 albumin genes per rat haploid genome. The number of AFP genes obtained in similar titration experiments was approximately 2--3. This was true whether rat liver DNA or hepatoma 7777 DNA was used in the reassociation experiments. When high molecular weight DNA preparations from both these tissues were digested with the restriction endonuclease EcoRI and the fragments were transferred to a nitrocellulose filter, the albumin and AFP [32P]cDNA probes hybridized to different sets of DNA fragments. However, each probe gave the same hybridization pattern whether Buffalo rat liver DNA or hepatoma 7777 DNA was utilized.

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Selected References

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  1. Abelev G. I. Alpha-fetoprotein as a marker of embryo-specific differentiations in normal and tumor tissues. Transplant Rev. 1974;20(0):3–37. doi: 10.1111/j.1600-065x.1974.tb00139.x. [DOI] [PubMed] [Google Scholar]
  2. Abelev G. I. Alpha-fetoprotein in ontogenesis and its association with malignant tumors. Adv Cancer Res. 1971;14:295–358. doi: 10.1016/s0065-230x(08)60523-0. [DOI] [PubMed] [Google Scholar]
  3. Blin N., Stafford D. W. A general method for isolation of high molecular weight DNA from eukaryotes. Nucleic Acids Res. 1976 Sep;3(9):2303–2308. doi: 10.1093/nar/3.9.2303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Britten R. J., Graham D. E., Neufeld B. R. Analysis of repeating DNA sequences by reassociation. Methods Enzymol. 1974;29:363–418. doi: 10.1016/0076-6879(74)29033-5. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Higashi K., Adams H. R., Busch H. Base composition of RNA fractions of nuclei of Walker tumor isolated with the citric acid procedure. Cancer Res. 1966 Oct;26(10):2196–2201. [PubMed] [Google Scholar]
  7. 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]
  8. McDonell M. W., Simon M. N., Studier F. W. Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol. 1977 Feb 15;110(1):119–146. doi: 10.1016/s0022-2836(77)80102-2. [DOI] [PubMed] [Google Scholar]
  9. Muto M. Estimation of gene reiteration from hybridization kinetics in moderate deoxyribonucleic acid excess. Biochem J. 1977 Jul 1;165(1):19–25. doi: 10.1042/bj1650019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Pearson W. R., Davidson E. H., Britten R. J. A program for least squares analysis of reassociation and hybridization data. Nucleic Acids Res. 1977 Jun;4(6):1727–1737. doi: 10.1093/nar/4.6.1727. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Pearson W. R., Wu J. R., Bonner J. Analysis of rat repetitive DNA sequences. Biochemistry. 1978 Jan 10;17(1):51–59. doi: 10.1021/bi00594a008. [DOI] [PubMed] [Google Scholar]
  12. Ruoslahti E., Pihko H., Seppälä M. Alpha-fetoprotein: immunochemical purification and chemical properties. Expression in normal state and in malignant and non-malignant liver disease. Transplant Rev. 1974;20(0):38–60. doi: 10.1111/j.1600-065x.1974.tb00140.x. [DOI] [PubMed] [Google Scholar]
  13. Savage M. J., Sala-Trepat J. M., Bonner J. Measurement of the complexity and diversity of poly(adenylic acid) containing messenger RNA from rat liver. Biochemistry. 1978 Feb 7;17(3):462–467. doi: 10.1021/bi00596a014. [DOI] [PubMed] [Google Scholar]
  14. Schreiber G., Rotermund H. M., Maeno H., Weigand K., Lesch R. The proption of the incorporation of leucine into albumin to that into totaprotein in rat liver and hepatoma Morris 5123 TC. Eur J Biochem. 1969 Sep;10(2):355–361. doi: 10.1111/j.1432-1033.1969.tb00698.x. [DOI] [PubMed] [Google Scholar]
  15. Sell S., Becker F. F., Leffert H. L., Watabe L. Expression of an oncodevelopmental gene product (alpha-fetoprotein) during fetal development and adult oncogenesis. Cancer Res. 1976 Nov;36(11 Pt 2):4239–4249. [PubMed] [Google Scholar]
  16. Sell S., Becker F. F. alpha-Fetoprotein. J Natl Cancer Inst. 1978 Jan;60(1):19–26. doi: 10.1093/jnci/60.1.19. [DOI] [PubMed] [Google Scholar]
  17. Sell S., Wepsic H. T., Nickel R., Nichols M. Rat alpha1 fetoprotein. IV. Effect of growth and surgical removal of Morris hepatoma 7777 on the serum alpha1F concentration of Buffalo rats. J Natl Cancer Inst. 1974 Jan;52(1):133–137. doi: 10.1093/jnci/52.1.133. [DOI] [PubMed] [Google Scholar]
  18. Sharp P. A., Sugden B., Sambrook J. Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose--ethidium bromide electrophoresis. Biochemistry. 1973 Jul 31;12(16):3055–3063. doi: 10.1021/bi00740a018. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Tse T. P., Morris H. P., Taylor J. M. Molecular basis of reduced albumin synthesis in Morris hepatoma 7777. Biochemistry. 1978 Jul 25;17(15):3121–3128. doi: 10.1021/bi00608a028. [DOI] [PubMed] [Google Scholar]
  21. Van Furth R., Adinolfi M. In vitro synthesis of the foetal alpha 1-globulin in man. Nature. 1969 Jun 28;222(5200):1296–1299. doi: 10.1038/2221296a0. [DOI] [PubMed] [Google Scholar]

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