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(A) Genetic homology of POWV strains LB and LI9 with differences indicated by white bars. (B) LB genomes (∼11 kb) were cloned into 5 overlapping fragments and purified <t>Phusion</t> amplified fragments were used develop reverse genetics. (C) POWV-LB fragments and UTR linkers, containing an HDVr, SV40pA, and CMVd2 promoter, were CPER circularized prior to transfection. (D) Immunoperoxidase staining of recLB infected VeroE6 cells 2-4 dpi. (E) Immunoperoxidase staining of WT LB or recLB infected VeroE6 cell foci (MOI, 0.1) 4 dpi.
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(A) Genetic homology of POWV strains LB and LI9 with differences indicated by white bars. (B) LB genomes (∼11 kb) were cloned into 5 overlapping fragments and purified Phusion amplified fragments were used develop reverse genetics. (C) POWV-LB fragments and UTR linkers, containing an HDVr, SV40pA, and CMVd2 promoter, were CPER circularized prior to transfection. (D) Immunoperoxidase staining of recLB infected VeroE6 cells 2-4 dpi. (E) Immunoperoxidase staining of WT LB or recLB infected VeroE6 cell foci (MOI, 0.1) 4 dpi.

Journal: bioRxiv

Article Title: Powassan Virus LB Neurovirulence and Lethality is Determined by Envelope Protein Domain III Residues

doi: 10.64898/2026.03.26.714546

Figure Lengend Snippet: (A) Genetic homology of POWV strains LB and LI9 with differences indicated by white bars. (B) LB genomes (∼11 kb) were cloned into 5 overlapping fragments and purified Phusion amplified fragments were used develop reverse genetics. (C) POWV-LB fragments and UTR linkers, containing an HDVr, SV40pA, and CMVd2 promoter, were CPER circularized prior to transfection. (D) Immunoperoxidase staining of recLB infected VeroE6 cells 2-4 dpi. (E) Immunoperoxidase staining of WT LB or recLB infected VeroE6 cell foci (MOI, 0.1) 4 dpi.

Article Snippet: DNA fragments 1-5 and linker (0.09 pmol each) were CPER amplified in a 25-μL reaction containing 200 μM of dNTPs, Phusion polymerase GC reaction buffer, and 0.5 μL Phusion polymerase (New England Biolabs).

Techniques: Clone Assay, Purification, Amplification, Transfection, Immunoperoxidase Staining, Infection