Ga. Smith et Lw. Enquist, A self-recombining bacterial artificial chromosome and its application foranalysis of herpesvirus pathogenesis, P NAS US, 97(9), 2000, pp. 4873-4878
Citations number
25
Categorie Soggetti
Multidisciplinary
Journal title
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
A self-recombining bacterial artificial chromosome (BAC) containing the 142
-kb pseudorabies virus genome was constructed such that the viral genome is
rapidly excised from the BAC vector backbone on delivery into mammalian ce
lls. The recombination is mediated by loxP sites in the plasmid and Cre rec
ombinase encoded within the BAC vector. A synthetic intron inserted in the
middle of the cre ORF completely inhibits recombination in Escherichia coli
, but is spliced out after delivery of the plasmid into mammalian cells. Re
combination is efficient. and pure virus lacking the BAC vector backbone is
immediately isolated from transfected mammalian cells without the need of
serial passage or plaque purification.