Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range.

MethodsX

National Microbiology Laboratory, the Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, Canada.

Published: May 2020

Vaccinia virus is capable of replicating in a wide range of vertebrate animal cells. However, deletion of the two virus host range genes, E3L and K3L, would render replication of the virus abortive in all the cell types tested, except the cells with defective PKR and RNase L activity. By expressing different poxvirus K3 ortholog proteins in the E3L and K3L double knockout vaccinia virus, we can construct a mutant vaccinia virus with modified host range. Here, using poxvirus K3 ortholog as a positive selection marker, we developed a proof-of-concept protocol to construct recombinant vaccinia viruses expressing foreign proteins. Such a recombinant virus has a modified host range in comparison to wild-type vaccinia virus. This protocol offers the following advantages:➢Cheap: no additional selection reagent is required.➢Highly efficient: nearly all recombinant virus plaques picked would be the stable recombinant virus expressing the protein of interest.➢Rapid: starting from transfection with the recombinant DNA vector, a purified recombinant virus expressing the protein of interest could be obtained within a week.

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7264055PMC
http://dx.doi.org/10.1016/j.mex.2020.100918DOI Listing

Publication Analysis

Top Keywords

host range
16
vaccinia virus
16
recombinant virus
16
poxvirus ortholog
12
modified host
12
virus
10
ortholog positive
8
positive selection
8
selection marker
8
recombinant vaccinia
8

Similar Publications

Want AI Summaries of new PubMed Abstracts delivered to your In-box?

Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!