Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
, 238 (2), 444-51

Construction and Characterization of Vaccinia Direct Ligation Vectors

Affiliations

Construction and Characterization of Vaccinia Direct Ligation Vectors

M Merchlinsky et al. Virology.

Abstract

Poxvirus vectors are extensively used as expression vehicles for protein and antigen expression in eukaryotic cells. Customarily, the foreign DNA is introduced into the poxvirus genome by homologous recombination. An alternative method using direct ligation vectors has been used to efficiently construct chimeric genomes in situations not readily amenable for homologous recombination. We describe the construction and characterization of a new set of direct ligation vectors designed to be universally applicable for the generation of chimeric vaccinia genomes. These vectors contain the pair of unique restriction sites NotI and ApaI to eliminate religation of poxvirus arms and fix the orientation of the insert DNA behind strongly expressing constitutive vaccinia promoters. The insertion cassette has been placed at the beginning of the thymidine kinase gene in vaccinia to use drug selection in the isolation of recombinants. These viruses provide a set of universally applicable direct ligation poxvirus cloning vectors, extending the utility of poxvirus vectors for construction and expression of complex libraries.

Similar articles

See all similar articles

Cited by 6 articles

See all "Cited by" articles

Publication types

LinkOut - more resources

Feedback