An improved cosmid vector for the cloning of equine herpesvirus DNA.
We have modified the commercial cosmid vector, triple helix vector (THV), such that I-Sce-I restriction endonuclease sites flank the cloning site. I-Sce-I is a rare-cutting endonuclease which recognizes an 18-bp sequence. It does not restrict the genome of either of the equine herpesvirus 1 or 4 (EHV-1 and EHV-4) strains we have cosmid cloned. Thus, cosmid-cloned EHV fragments can be excised intact from the vector by I-Sce-I digestion, facilitating production of large overlapping EHV fragments for use in transfections to produce recombinant virus.
Nicolson
L
L
Department of Veterinary Pathology, University of Glasgow Veterinary School, UK.
Rafferty
E L
EL
Brawley
A
A
Onions
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DE
D001483:Base Sequence; D003001:Cloning, Molecular / Q000379:methods*; D003360:Cosmids; D004279:DNA, Viral / Q000235:genetics*; D005822:Genetic Vectors; D004861:Herpesvirus 1, Equid / Q000235:genetics*; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D015183:Restriction Mapping; D018140:Varicellovirus / Q000235:genetics*
D004279:DNA, Viral; D009838:Oligodeoxyribonucleotides