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J Virol. 1971 March; 7(3): 343-351
Copyright © 1971 American Society for Microbiology. All Rights Reserved.

Studies of Nondefective Adenovirus 2-Simian Virus 40 Hybrid Viruses II. Relationship of Adenovirus 2 Deoxyribonucleic Acid and Simian Virus 40 Deoxyribonucleic Acid in the Ad2⁺ND₁ Genome

^a Medicine Branch, National Cancer Institute, and Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20014

ABSTRACT

A nondefective adenovirus 2 (Ad2)-simian virus 40 (SV40) hybrid virus, Ad2⁺ND₁, has been plaque-isolated from an Ad2-SV40 hybrid population. This virus, unlike the defective Ad-SV40 hybrid populations previously described, replicates without the aid of nonhybrid adenovirus helper. Consequently, the hybrid virus deoxyribonucleic acid (DNA) can be obtained free of nonhybrid adenovirus DNA. The DNA of the Ad2⁺ND₁ virus was shown by ribonucleic acid (RNA)-DNA hybridization to consist of nucleotide sequences complementary to Ad2- and SV40-specific RNA. Techniques of equilibrium density and rate zonal centrifugation were employed to demonstrate that these Ad2 and SV40 nucleotide sequences were linked together in the same DNA molecules by alkali-resistant bonds. Calibration curves were established relating the amount of tritium-labeled SV40-specific RNA (prepared in vitro or in vivo) bound to given amounts of SV40 DNA in a hybridization reaction, and these curves were employed to determine the equivalent amount of SV40 DNA in the Ad2⁺ND₁ molecule. From the results obtained, it was estimated that 1% of the Ad2⁺ND₁ DNA consists of SV40 nucleotide sequences.

¹ Present address: Research Division of Infectious Diseases, Children's Hospital Medical Center and Departments of Medicine and Bacteriology and Immunology, Harvard Medical School, Boston, Mass. 02115.

² Present address: Department of Medicine, University of Missouri, Columbia, Mo. 65201.