Comparative Studies on the Structure of Human Adenovirus Genomes 4, 7 and 21.
Annual progress rept. 1 Feb 80-2 Feb 81,
MARYLAND UNIV BALTIMORE DEPT OF BIOLOGICAL CHEMISTRY
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A sensitive method was developed to label the 5 termini of Ad DNA which was found to be applicable to Ad 7, Ad 4 and Ad 21 DNA due to the presence of a tyrosine-containing peptide covalently attached to these DNA molecules. Ad 7 vaccine strain was passaged in 293 cells and then grown in large amounts in suspension cultures of KB cells. The DNA was extracted and purified. Restriction enzyme analysis of vaccine and Greider Ad 7 strains revealed that the two strains gave identical cleavage patterns with 8 restriction enzymes. Ad 4 prototype strain ATCC have been passaged in 293 cells and then grown in large amounts in suspension cultures of human KB cells. The DNA from these virions have been extracted and purified. Ad 21 vaccine strain has been passaged in 293 cells and then the virus was grown in KB cell suspension cultures. We are currently in the process of isolating the DNA and analyzing its genomic organization with several restriction enzymes. We analyzed the efficiencies of different protocols for titering the live Ad vaccine strains Ad 4, Ad 7 and Ad 21 present in enteric coated tablets.