Cloning and physical mapping of enteric adenoviruses (candidate types 40 and 41)

Abstract

We have studied the DNAs of fastidious enteric adenoviruses recovered from the stools of infants with gastroenteritis. By endonuclease analysis, the strains examined represent candidate adenovirus types 40 and 41, which are thought to comprise new adenovirus subgroups F and G. Cloning of DNA from representative enteric adenovirus isolates, together with hybridization and subcleavage analysis, permitted the mapping of restriction enzyme cleavage sites. Although the restriction profiles are different for the two strains, they appear to have several cleavage sites in common. Cross hybridization studies show considerable homology between the subgroup F and G strains but much less homology to adenovirus 2. In addition, regions on both ends of enteric adenovirus genomes (map units, 2.9 to 11.3 and 75 to 100) possess little or no homology to adenovirus 2. Restriction enzyme digests reveal submolar fragments that map to the terminal regions of the genome. Electron micrographic studies of denatured and renatured DNA strands suggest that the submolar fragments may derive from cleavage of defective molecules. Inverted terminal repeat sequences were shown to comprise 0 to 3.2% of the length of complete (_22 megadaltons) enteric adenovirus DNA molecules but 4 to 69% of incomplete-length (<22-megadalton) molecules.

Specific detection and typing of adenovirus types 40 and 41 in stool specimens by dot-blot hybridization

“…these adenoviruses is reinforced by the DNA hybridization studies presented in this paper and elsewhere (1,20,24). The genomes of Ad4O and Ad4l are more closely related to each other than to any other human adenoviruses, and their initial provisional classification into two subgroups (F and G) (22,25) has been superseded by a single subgroup concept…”

“…3). By comparison with the Ad4O maps published by Takiff et al (20), the Ad4O-specific insert of plasmid N26 would span map units 68 through 73 (Fig. 4).…”

Enteric Adenovirus Type 41 Isolates: Cloning, Physical Maps and Diversity in Restriction Enzyme Cleavage Pattern

“…Ad41 shares many antigenic determinants with Ad40 (4), which suggests a close evolutionary relationship between these adenoviruses (1,22,32). However, our results revealed that Ad41 frequently showed genome variants, but not Ad40 in Japan.…”

“…Thus, it is of interest to compare these enteric Ad genome DNAs to each other and to other Ads. Viral DNAs from some laboratory propagated strains of Ad40 (32) and Ad41 (1,22), and from an Ad40 clinical strain isolated directly from stool specimen were cloned and characterized (9, 10, Demura et al in preparation).…”

Viruses

“…Serotypes 1 to 39 fall into subgroups A to D, except for serotype 4, whose pattern does not fit and which has thus been placed into subgroup E. The fastidious serotypes do not fit subgroups A to E, and are placed into separate subgroups (Wadell, 1984). Using restriction endonuclease analysis, serotypes 40 and 41 appear to be different and thus are given separate subgroups, although they appear to have several cleavage sites in common (de Jong et al, 1983;Uhnoo et al, 1983;Takiff et al, 1984). However, they have biological similarities and are indistinguishable by haemagglutination inhibition (Kidd, 1984;Christensen, 1989).…”