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α-fetoprotein and Albumin Genes of Rats: No Evidence for Amplification-Deletion or Rearrangement in Rat Liver Carcinogenesis

Sala-Trepat, José M. and Sargent, Thomas D. and Sell, Stewart and Bonner, James (1979) α-fetoprotein and Albumin Genes of Rats: No Evidence for Amplification-Deletion or Rearrangement in Rat Liver Carcinogenesis. Proceedings of the National Academy of Sciences of the United States of America, 76 (2). pp. 695-699. ISSN 0027-8424.

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Full-length radiolabeled albumin and alpha --fetoprotein (AFP) cDNAs were synthesized from pure albumin and AFP 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 mRNAs 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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Additional Information:Copyright © 1979 by the National Academy of Sciences Contributed by James F. Bonner, November 20, 1978 We thank Dr. Harold Morris, who graciously furnished the tumor-bearing rats. We are indebted to Dr. J. Beard for providing us with reverse transcriptase of avian myeloblastosis virus as part of the joint National Institutes of Health-National Cancer Institute program. We also thank Dr. J. Dever and K. Thomas for help in the early part of this work, R. F. Murphy for help in the computer analysis of the hybridization and reassociation data, and James Posakony for helpful discussion. The skillful technical assistance of John Rising is gratefully acknowledged. This work was supported by U.S. Public Health Service Grant GM-13762, by National Cancer Institute Grant CA-22227, and by the French Research Council (Centre National de la Recherche Scientifique). The publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact.
Subject Keywords:gene frequency; gene regulation; cDNA; DNA hybridization; "blot" hybridization
Issue or Number:2
Record Number:CaltechAUTHORS:SALpnas79
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Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:6778
Deposited By: Archive Administrator
Deposited On:21 Dec 2006
Last Modified:02 Oct 2019 23:35

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