Summary
Restriction endonuclease maps were constructed for the genome of a porcine adenovirus (PAV), NADC-1, which was isolated in 1972 from an adult swine. Genomic DNA libraries of NADC-1 Bam HI, Eco RI/Bam HI, and Sph I fragments were cloned into pUC-18. Using the cloned NADC-1 Bam HI and Eco RI/Bam HI fragments as probes, Southern blot hybridizations were performed to human adenovirus 2 (Ad-2) restriction fragments to determine the left-to-right orientation of the Bam HI and Eco RI/Bam HI fragments. Genomic NADC-1 DNA was cleaved with ten restriction endonucleases (RE). Using cloned NADC-1 genomic fragments as probes in Southern blot hybridizations, an RE site map was constructed. Nucleotide sequencing of four clones confirmed several RE sites. The size of the NADC-1 genome was determined to be approximately 32 kbp. The size of Hind III, Xba I, Sma I, Eco RI, Bam HI, Bgl II, Pst I, and Sph I RE fragments from NADC-1 was compared to those from the reference strain of PAV serotype 4 (F 618), and to two recent isolates, NADC-2 and NADC-3. For all restriction enzymes examined, the sizes of the NADC-1 fragments were identical to PAV-4, NADC-2, and NADC-3 fragments, indicating that the NADC-1 isolate is very closely related, if not identical, to PAV-4 and two recent isolates. Southern blot hybridizations also indicated that NADC-1, NADC-2, NADC-3, and PAV-4 are very similar and revealed regions of sequence similarity between NADC-1 and human Ad-2 and human Ad-5.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Benkö M, Harrach B, D'Halluin J (1990) Molecular cloning and physical mapping of the DNA of bovine adenovirus serotype 4; study of the DNA homology among bovine, human, and porcine adenoviruses. J Gen Virol 71: 465–469
Culpepper JH, Angermüller SA, Sayavedra-Soto LA (1990) Mini-Prep in ten minutes. BioTechniques 8: 72–173
Dea S, El Azhary MASY (1984) Prevalence of antibodies to porcine adenovirus in swine by indirect fluorescent antibody test. Am J Vet Res 45: 2109–2112
Derbyshire JB (1992) Adenovirus. In: Leman AD, Straw BE, Mengeling WL, D'Allaire S, Taylor DJ (eds) Diseases of swine, 7th edn. Iowa State University Press, Ames, pp 225–227
Dragulev BP, Sira S, Abouhaidar MG, Campbell JB (1991) Sequence analysis of putative E 3 and fiber genomic regions of two strains of canine adenovirus type 1. Virology 183: 298–305
Freshney R (1987) Culture of animal cells: a manual of basic techniques, 2nd edn. Liss, New York
Garwes DJ, Xuan H (1989) Genome typing of three serotypes of porcine adenovirus. Intervirology 30: 234–236
Grunhaus A, Horwitz MS (1992) Adenoviruses as cloning vectors. Semin Virol 3: 237–252
Haig DA, Clarke MC, Pereira MS (1964) Isolation of an adenovirus from a pig. J Comp Pathol 74: 81–84
Hanahan D (1985) Techniques for transformation of E. coli. In: Glover DM (ed) DNA cloning: a practical approach. IRL Press, Oxford, pp 109–135
Hanahan D, Gluzman Y (1984) Rescue of functional replication origins from embedded configurations in a plasmic carrying the adenovirus genome. Mol Cell Biol 4: 302–309
Hirt B (1967) Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol 26: 365–369
Horwitz MS (1991) Adenoviridae and their replication. In: Fields BN, Knipe DM (eds) Fundamentals of virology, 2nd edn. Raven Press, New York, pp 771–813
Kasza L (1966) Isolation of an adenovirus from the brain of a pig. Am J Vet Res 27: 751–758
Mittal SK, Prevec L, Babiuk LA, Graham FL (1992) Sequence analysis of bovine adenovirus type-3 early region-3 and fibre protein genes. J Gen Virol 73: 3295–3300
Rekosh DMK, Russell WC, Bellet AJD, Robinson AJ (1977) Identification of a protein linked to the ends of adenovirus DNA. Cell 11: 283–295
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor
Sanger F, Nicklen S, Coulson AR (1977) DNA sequencing with chain terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5464
Sheppard M, Drysdale SM, Studdert MJ (1992) Restriction enzyme maps for equine adenovirus-1 genome. Vet Microbiol 31: 343–349
Shinagawa M, Matsuda A, Ishiyama T, Goto H, Sato G (1983) A rapid and simple method for preparation of adenovirus DNA from infected cells. Microbiol Immunol 27: 817–822
Southern EM (1975) Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol 98: 503–517
Struhl K (1985) A rapid method for creating recombinant DNA molecules. BioTechniques 3: 452–453
Tabor S, Richardson CC (1987) DNA sequence analysis with a modified bacteriophage T 7 DNA polymerase. Proc Natl Acad Sci USA 84: 4767–4771
Wold WSM, Gooding LR (1991) Region E 3 of adenovirus: a cassette of genes involved in host immunosurveillance and virus-cell interactions. Virology 184: 1–8
Yanisch-Perron C, Vieira J, Messing J (1985) Improved M 13 phage cloning vectors and host strains: nucleotide sequences of the M 13 mp 18 and pUC 19 vectors. Gene 33: 103–119
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kleiboeker, S.B., Seal, B.S. & Mengeling, W.L. Genomic cloning and restriction site mapping of a porcine adenovirus isolate: demonstration of genomic stability in porcine adenovirus. Archives of Virology 133, 357–368 (1993). https://doi.org/10.1007/BF01313775
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF01313775