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1
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pTAV-2 plasmid
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Used as the source plasmid for mutagenesis and generation of wild-type virus stocks.
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7996133
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View Related Publication
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2
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pTAV-p plasmid
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Used to generate a virus mutant intended to replace the RGD motif with RGE.
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3
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pTAV-d plasmid
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Used to generate a virus mutant intended to delete the RGD motif.
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4
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M13mpl8 vector
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Used for cloning the 2.9 kb HindIII/SstI fragment of the capsid protein sequence for site-directed mutagenesis.
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5
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Amersham mutagenesis kit
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Used for performing site-directed mutagenesis following the supplier's instructions.
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6
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Baculovirus-expressed VP3 protein
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Used for protein sequencing to confirm the corrected amino acid sequence.
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7
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Endoproteinase lys C
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Used for digestion of baculovirus-expressed VP3 protein during protein sequencing.
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8
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1% NP40, 150 mM-NaCl, 20 mM-Tris-HCl (pH 8), 10 mM-DTT, 5 mM-EDTA
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Lysis buffer for cell lysis followed by sedimentation during protein sequencing.
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9
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8 M-urea, 20 mM-Tris-HCl (pH 7.5)
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Used to resuspend the pellet after sedimentation in protein sequencing.
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10
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SDS-PAGE
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Used for separating proteins during gel electrophoresis.
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11
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Immobilon-P membrane (Millipore)
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Used for transferring proteins after SDS-PAGE.
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12
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Ponceau S
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Used for staining the transferred proteins to visualize VP3 position.
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13
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200 mM-Tris-HCl (pH 8), 0.2% Tween-20, 5 mM-DTT
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Incubation buffer for membrane pieces containing VP3.
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14
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5 M-guanidinium-HCl, 200 mM-Tris-HCl (pH 8), 5 mM-DTT
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Buffer for digesting membrane-bound polypeptides with endoproteinase lys C.
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15
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Trifluoroacetic acid
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Added to digest peptides for purification.
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16
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Protein sequencer (Applied Biosystems, model 477A)
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Used for sequencing the purified peptides.
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17
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Antibodies
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Various antibodies were used for immunoblotting and immunofluorescence analysis.
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18
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Peroxidase-conjugated secondary antibodies
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Used for visualizing capsid proteins in immunoblots via enhanced chemiluminescence detection.
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19
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Hirt extraction reagents
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Used to prepare extracts for DNA replication analysis.
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20
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EVL Western blotting
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Used for detecting sedimentable capsid proteins after high-speed centrifugation.
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21
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Rhodamine-conjugated secondary antibodies
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Used for visualizing bound primary antibodies in immunofluorescence staining.
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22
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Fluorescein isothiocyanate-conjugated secondary antibodies
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Used for visualizing bound primary antibodies in immunofluorescence double staining.
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23
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Radiolabelled NdeI/XbaI fragment
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Used as a probe in dot-blot hybridization.
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