Restriction endonuclease analysis of alcelaphine herpesvirus 1 DNA and molecular cloning of virus genomic DNA for potential diagnostic use.
Alcelaphine herpesvirus 1 (AHV-1) genomic DNA was analyzed using restriction enzymes having recognition sequences both low in guanine-cytosine content (BamHI, KpnI, HindIII) and high in guanine-cytosine content (SmaI, AvaI, ApaI). The results from the restriction enzyme analyses along with exonuclease treatment demonstrated that the termini of AHV-1 DNA are likely composed of polyrepetitive sequences high in guanine-cytosine content similar to those found in Herpesvirus saimiri DNA. Cleaving AHV-1 DNA with the restriction enzyme SmaI produced polyrepetitive sequences that appeared as low molecular weight supermolar bands less than 1 kilobase pairs (kb) in size. Analysis of AHV-1 DNA cleaved with HindIII indicated that the polyrepetitive sequences are approximately 29.5 kb in size. The total molecular weight for AHV-1 DNA was determined to be approximately 118 kb. Seventy-five percent of the AHV-1 genome was cloned into the plasmids pAT153 and pBR322. Cloned DNA fragments representing unique sequences of the AHV-1 genome hybridized with AHV-1 genomic DNA but showed no appreciable hybridization with AHV-2 DNA or bovine herpesvirus 1, 2, or 4 DNA.
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Center for Reproduction of Endangered Species, Zoological Society of San Diego, CA 92112.
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D000818:Animals; D000866:Antelopes; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000032:analysis* / Q000235:genetics; D003720:Densitometry; D004587:Electrophoresis, Agar Gel; D005822:Genetic Vectors; D006564:Herpesviridae / Q000235:genetics*; D006566:Herpesviridae Infections / Q000175:diagnosis / Q000662:veterinary*; D008304:Malignant Catarrh / Q000175:diagnosis*; D009693:Nucleic Acid Hybridization; D010957:Plasmids; D015183:Restriction Mapping
D004279:DNA, Viral