Molecular engineering of the herpes simplex virus genome: Insertion of a second L-S junction into the genome causes additional genome inversions

摘要: We have developed a technique for the insertion of any DNA fragment into herpes simplex virus (HSV) genome at specific sites. This was used to resolve problem concerning isomerization HSV genome. Briefly, consists four isomers differing in orientation two covalently linked components, L and S, relative each other. Each component unique sequences flanked by inverted repeats. To determine whether is result generalized recombinatin between homologous reiterated repeats or site-specific recombination, we constructed plasmids which fragments derived from various regions viral were inserted both orientations thymidine kinase gene, rendering it nonfunctional. The then recombined genome, recombinants selected their kinase-deficient phenotype. these recombination highly efficient that all clones isolated contained expected location. only promoted additional inversions those spanning junction S components. Furthermore, analysis formed indicates occur when are relation termini authentic junction.