Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range
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...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2020-01-01
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| Series: | MethodsX |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2215016120301370 |
| _version_ | 1818720182041837568 |
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| author | Jingxin Cao Christine Layne Jessie Varga Yvon Deschambault |
| author_facet | Jingxin Cao Christine Layne Jessie Varga Yvon Deschambault |
| author_sort | Jingxin Cao |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T20:18:46Z |
| format | Article |
| id | doaj.art-554e741b6a2845a4ba475a2102a3016c |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-17T20:18:46Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-554e741b6a2845a4ba475a2102a3016c2022-12-21T21:34:00ZengElsevierMethodsX2215-01612020-01-017100918Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host rangeJingxin Cao0Christine Layne1Jessie Varga2Yvon Deschambault3National Microbiology Laboratory, the Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, Canada; Department of Medical Microbiology, University of Manitoba, 543-745 Bannatyne Avenue, Winnipeg, Manitoba R3E 0J9, Canada; Corresponding author.National Microbiology Laboratory, the Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, CanadaNational Microbiology Laboratory, the Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, CanadaNational Microbiology Laboratory, the Public Health Agency of Canada, 1015 Arlington Street, Winnipeg, Manitoba R3E 3R2, CanadaVaccinia 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.http://www.sciencedirect.com/science/article/pii/S2215016120301370PoxvirusVacciniaRecombinationHost rangek3L |
| spellingShingle | Jingxin Cao Christine Layne Jessie Varga Yvon Deschambault Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range MethodsX Poxvirus Vaccinia Recombination Host range k3L |
| title | Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| title_full | Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| title_fullStr | Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| title_full_unstemmed | Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| title_short | Application of poxvirus K3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| title_sort | application of poxvirus k3 ortholog as a positive selection marker for constructing recombinant vaccinia viruses with modified host range |
| topic | Poxvirus Vaccinia Recombination Host range k3L |
| url | http://www.sciencedirect.com/science/article/pii/S2215016120301370 |
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