Comparative Studies on the Structure of Human Adenovirus Genomes 4, 7 and 21.
Annual progress rept. 1 Jul 79-20 Jan 80,
MARYLAND UNIV BALTIMORE DEPT OF BIOLOGICAL CHEMISTRY
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Adenovirus Type 7 Greider Strain was grown in human KB cells. The virions were purified by CsCl equilibrium centrifugation, the DNA was extracted and purified. The viral genome was analyzed by restriction endonucleases EcoRl, BamHI, SmaI, HpaI, BelI, BstEII and KpnI. The restriction endonuclease patterns of the viral DNA was compared with those of three Ad 7 subgroups published by Wadell and Varsanyi 14. Patterns we obtained by cleaving Ad 7 with SalI, HpaI, Bam Hi and SmaI were identical to those obtained by cleaving Ad 7 with SalI, HpaI, Bam Hi and SmaI were identical to those obtained with strain 1058 and strain 55142 Ad 7 vaccine strain reported by Wadell and Varsanyi 14. However, EcoRI pattern of Ad 7 Greider was different from strain 1058 and 55142 in that one site mapped at 85.4 units from the left end of the DNA is missing. DNA sequence at the inverted terminal repetition of Ad 7 DNA was analyzed in order to compare Ad 7 Greider with Ad 3 DNA, another member of weakly oncogenic group B. Ad 2 add Ad 5 member of non-oncogenic group C and with Ad 12, member of highly oncogenic group A of human adenoviruses. Our results indicate that the length of the inverted terminal repetition of Ad 7 Greider is identical to Ad 3 but different from Ad 2 or Ad 5 and Ad 12 or Ad 18. The nucleotide sequence homology in this unique region between Ad 7 and Ad 3 is 95. We purified the Ad 7 DNA-protein complex to establish conditions for DNA-transfection in Human Embryonic Kidney cells in vitro.
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