Construction of a novel set of transfer vectors to study vaccinia virus replication and foreign gene expression.
Vaccinia virus (VV) is a useful expression vector for many laboratory applications. To date, approximately 60% ( approximately 120) of the VV genes remain uncharacterized. The thought of smallpox being used as a biological weapon has gained attention. In light of this, it is imperative that we continue to study the basic replication of VV, a poxvirus that is closely related to smallpox. A set of plasmid vectors were constructed to generate gene deletions (pZIPPY-NEO/GUS) in VV or for foreign gene expression (pBR-EXPRESS). The vectors contain the Escherichia coli neomycin resistance (neo) and beta-glucuronidase (gusA) genes as selectable markers to facilitate isolation of recombinant viruses. These are the first transfer vectors to use a neo/gusA selection system. We used these vectors to successfully generate a recombinant D9R deletion mutant of VV and to express E. coli lacZ gene. Results indicate that both vectors are highly suited for their designed purpose.
Dvoracek
Barbara
B
Department of Biology and Microbiology, University of Wisconsin-Oshkosh, Oshkosh, WI, USA.
Shors
Teri
T
D000818:Animals; D001483:Base Sequence; D002478:Cells, Cultured; D004251:DNA Transposable Elements; D021901:DNA, Intergenic; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000235:genetics; D005966:Glucuronidase / Q000235:genetics; D019868:Kanamycin Kinase / Q000235:genetics; D007668:Kidney / Q000166:cytology / Q000821:virology; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics; D014616:Vaccinia virus / Q000235:genetics*; D014779:Virus Replication / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D004251:DNA Transposable Elements; D021901:DNA, Intergenic; D011994:Recombinant Proteins; D019868:Kanamycin Kinase; D001616:beta-Galactosidase; D005966:Glucuronidase