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Manuel V Borca - One of the best experts on this subject based on the ideXlab platform.

  • the l83l orf of african swine fever virus strain georgia encodes for a non Essential Gene that interacts with the host protein il 1β
    Virus Research, 2018
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Marialexia Alfano, Elizabeth Ramirezmedina, Benjamin A Clark, Elizabeth A Vuono, Keith A Berggren, Lucas B Carey, Juergen A Richt
    Abstract:

    African swine fever virus (ASFV) causes a contagious and frequently lethal disease of pigs causing significant economic consequences to the swine industry. The ASFV genome encodes for more than 150 Genes, but only a few of them have been studied in detail. Here we report the characterization of open reading frame L83L which encodes a highly conserved protein across all ASFV isolates. A recombinant ASFV harboring a HA tagged L83L protein was developed (ASFV-G-L83L-HA) and used to demonstrate that L83L is a transiently expressed early virus protein. A recombinant ASFV lacking the L83L Gene (ASFV-G-ΔL83L) was developed from the highly virulent field isolate Georgia2007 (ASFV-G) and was used to show that L83L is a non-Essential Gene. ASFV-G-ΔL83L had similar replication in primary swine macrophage cells when compared to its parental virus ASFV-G. Analysis of host-protein interactions for L83L identified IL-1β as its host ligand. Experimental infection of domestic pigs showed that ASFV-G-ΔL83L is as virulent as the parental virus ASFV-G.

  • the ep152r orf of african swine fever virus strain georgia encodes for an Essential Gene that interacts with host protein bag6
    Virus Research, 2016
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Devendra K Rai, Brenton Sanford, Marialexia Alfano, Jolene Carlson, Paul A Azzinaro, Covadonga Alonso, Douglas P Gladue
    Abstract:

    African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The viral genome encodes for more than 150 Genes, and only a select few of these Genes have been studied in some detail. Here we report the characterization of open reading frame Ep152R that has a predicted complement control module/SCR domain. This domain is found in Vaccinia virus proteins that are involved in blocking the immune response during viral infection. A recombinant ASFV harboring a HA tagged version of the Ep152R protein was developed (ASFV-G-Ep152R-HA) and used to demonstrate that Ep152R is an early virus protein. Attempts to construct recombinant viruses having a deleted Ep152R Gene were consistently unsuccessful indicating that Ep152R is an Essential Gene. Interestingly, analysis of host-protein interactions for Ep152R using a yeast two-hybrid screen, identified BAG6, a protein previously identified as being required for ASFV replication. Furthermore, fluorescent microscopy analysis confirms that Ep152R-BAG6 interaction actually occurs in cells infected with ASFV.

Stephen A Osmani - One of the best experts on this subject based on the ideXlab platform.

  • identification and analysis of Essential aspergillus nidulans Genes using the heterokaryon rescue technique
    Nature Protocols, 2006
    Co-Authors: Aysha H Osmani, Berl R Oakley, Stephen A Osmani
    Abstract:

    In the heterokaryon rescue technique, Gene deletions are carried out using the pyrG nutritional marker to replace the coding region of target Genes via homologous recombination in Aspergillus nidulans. If an Essential Gene is deleted, the null allele is maintained in spontaneously Generated heterokaryons that consist of two Genetically distinct types of nuclei. One nuclear type has the Essential Gene deleted but has a functional pyrG allele (pyrG+). The other has the wild-type allele of the Essential Gene but lacks a functional pyrG allele (pyrG−). Thus, a simple growth test applied to the uninucleate asexual spores formed from primary transformants can identify deletions of Genes that are non-Essential from those that are Essential and can only be propagated by heterokaryon rescue. The growth tests also enable the phenotype of the null allele to be defined. Diagnostic PCR can be used to confirm deletions at the molecular level. This technique is suitable for large-scale Gene-deletion programs and can be completed within 3 weeks. Note: In the version of this article initially published, the black ball in Figure 2c was incorrectly described as representing a pyrG–, GeneX+ nuclei. This ball represents pyrG+, GeneX–. The error has been corrected in all versions of the article.

  • identification and analysis of Essential aspergillus nidulans Genes using the heterokaryon rescue technique
    Nature Protocols, 2006
    Co-Authors: Aysha H Osmani, Berl R Oakley, Stephen A Osmani
    Abstract:

    In the heterokaryon rescue technique, Gene deletions are carried out using the pyrG nutritional marker to replace the coding region of target Genes via homologous recombination in Aspergillus nidulans. If an Essential Gene is deleted, the null allele is maintained in spontaneously Generated heterokaryons that consist of two Genetically distinct types of nuclei. One nuclear type has the Essential Gene deleted but has a functional pyrG allele (pyrG+). The other has the wild-type allele of the Essential Gene but lacks a functional pyrG allele (pyrG-). Thus, a simple growth test applied to the uninucleate asexual spores formed from primary transformants can identify deletions of Genes that are non-Essential from those that are Essential and can only be propagated by heterokaryon rescue. The growth tests also enable the phenotype of the null allele to be defined. Diagnostic PCR can be used to confirm deletions at the molecular level. This technique is suitable for large-scale Gene-deletion programs and can be completed within 3 weeks.

Douglas P Gladue - One of the best experts on this subject based on the ideXlab platform.

  • the ep152r orf of african swine fever virus strain georgia encodes for an Essential Gene that interacts with host protein bag6
    Virus Research, 2016
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Devendra K Rai, Brenton Sanford, Marialexia Alfano, Jolene Carlson, Paul A Azzinaro, Covadonga Alonso, Douglas P Gladue
    Abstract:

    African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The viral genome encodes for more than 150 Genes, and only a select few of these Genes have been studied in some detail. Here we report the characterization of open reading frame Ep152R that has a predicted complement control module/SCR domain. This domain is found in Vaccinia virus proteins that are involved in blocking the immune response during viral infection. A recombinant ASFV harboring a HA tagged version of the Ep152R protein was developed (ASFV-G-Ep152R-HA) and used to demonstrate that Ep152R is an early virus protein. Attempts to construct recombinant viruses having a deleted Ep152R Gene were consistently unsuccessful indicating that Ep152R is an Essential Gene. Interestingly, analysis of host-protein interactions for Ep152R using a yeast two-hybrid screen, identified BAG6, a protein previously identified as being required for ASFV replication. Furthermore, fluorescent microscopy analysis confirms that Ep152R-BAG6 interaction actually occurs in cells infected with ASFV.

Vivian Odonnell - One of the best experts on this subject based on the ideXlab platform.

  • the l83l orf of african swine fever virus strain georgia encodes for a non Essential Gene that interacts with the host protein il 1β
    Virus Research, 2018
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Marialexia Alfano, Elizabeth Ramirezmedina, Benjamin A Clark, Elizabeth A Vuono, Keith A Berggren, Lucas B Carey, Juergen A Richt
    Abstract:

    African swine fever virus (ASFV) causes a contagious and frequently lethal disease of pigs causing significant economic consequences to the swine industry. The ASFV genome encodes for more than 150 Genes, but only a few of them have been studied in detail. Here we report the characterization of open reading frame L83L which encodes a highly conserved protein across all ASFV isolates. A recombinant ASFV harboring a HA tagged L83L protein was developed (ASFV-G-L83L-HA) and used to demonstrate that L83L is a transiently expressed early virus protein. A recombinant ASFV lacking the L83L Gene (ASFV-G-ΔL83L) was developed from the highly virulent field isolate Georgia2007 (ASFV-G) and was used to show that L83L is a non-Essential Gene. ASFV-G-ΔL83L had similar replication in primary swine macrophage cells when compared to its parental virus ASFV-G. Analysis of host-protein interactions for L83L identified IL-1β as its host ligand. Experimental infection of domestic pigs showed that ASFV-G-ΔL83L is as virulent as the parental virus ASFV-G.

  • the ep152r orf of african swine fever virus strain georgia encodes for an Essential Gene that interacts with host protein bag6
    Virus Research, 2016
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Devendra K Rai, Brenton Sanford, Marialexia Alfano, Jolene Carlson, Paul A Azzinaro, Covadonga Alonso, Douglas P Gladue
    Abstract:

    African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The viral genome encodes for more than 150 Genes, and only a select few of these Genes have been studied in some detail. Here we report the characterization of open reading frame Ep152R that has a predicted complement control module/SCR domain. This domain is found in Vaccinia virus proteins that are involved in blocking the immune response during viral infection. A recombinant ASFV harboring a HA tagged version of the Ep152R protein was developed (ASFV-G-Ep152R-HA) and used to demonstrate that Ep152R is an early virus protein. Attempts to construct recombinant viruses having a deleted Ep152R Gene were consistently unsuccessful indicating that Ep152R is an Essential Gene. Interestingly, analysis of host-protein interactions for Ep152R using a yeast two-hybrid screen, identified BAG6, a protein previously identified as being required for ASFV replication. Furthermore, fluorescent microscopy analysis confirms that Ep152R-BAG6 interaction actually occurs in cells infected with ASFV.

Lauren G Holinka - One of the best experts on this subject based on the ideXlab platform.

  • the l83l orf of african swine fever virus strain georgia encodes for a non Essential Gene that interacts with the host protein il 1β
    Virus Research, 2018
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Marialexia Alfano, Elizabeth Ramirezmedina, Benjamin A Clark, Elizabeth A Vuono, Keith A Berggren, Lucas B Carey, Juergen A Richt
    Abstract:

    African swine fever virus (ASFV) causes a contagious and frequently lethal disease of pigs causing significant economic consequences to the swine industry. The ASFV genome encodes for more than 150 Genes, but only a few of them have been studied in detail. Here we report the characterization of open reading frame L83L which encodes a highly conserved protein across all ASFV isolates. A recombinant ASFV harboring a HA tagged L83L protein was developed (ASFV-G-L83L-HA) and used to demonstrate that L83L is a transiently expressed early virus protein. A recombinant ASFV lacking the L83L Gene (ASFV-G-ΔL83L) was developed from the highly virulent field isolate Georgia2007 (ASFV-G) and was used to show that L83L is a non-Essential Gene. ASFV-G-ΔL83L had similar replication in primary swine macrophage cells when compared to its parental virus ASFV-G. Analysis of host-protein interactions for L83L identified IL-1β as its host ligand. Experimental infection of domestic pigs showed that ASFV-G-ΔL83L is as virulent as the parental virus ASFV-G.

  • the ep152r orf of african swine fever virus strain georgia encodes for an Essential Gene that interacts with host protein bag6
    Virus Research, 2016
    Co-Authors: Manuel V Borca, Vivian Odonnell, Lauren G Holinka, Devendra K Rai, Brenton Sanford, Marialexia Alfano, Jolene Carlson, Paul A Azzinaro, Covadonga Alonso, Douglas P Gladue
    Abstract:

    African swine fever virus (ASFV) is the etiological agent of a contagious and often lethal disease of domestic pigs that has significant economic consequences for the swine industry. The viral genome encodes for more than 150 Genes, and only a select few of these Genes have been studied in some detail. Here we report the characterization of open reading frame Ep152R that has a predicted complement control module/SCR domain. This domain is found in Vaccinia virus proteins that are involved in blocking the immune response during viral infection. A recombinant ASFV harboring a HA tagged version of the Ep152R protein was developed (ASFV-G-Ep152R-HA) and used to demonstrate that Ep152R is an early virus protein. Attempts to construct recombinant viruses having a deleted Ep152R Gene were consistently unsuccessful indicating that Ep152R is an Essential Gene. Interestingly, analysis of host-protein interactions for Ep152R using a yeast two-hybrid screen, identified BAG6, a protein previously identified as being required for ASFV replication. Furthermore, fluorescent microscopy analysis confirms that Ep152R-BAG6 interaction actually occurs in cells infected with ASFV.