Panoramic Parvovirus Strain Discovery Data Chain

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ParvoDB ID PV001334 GenBank Accession FJ560720
Subfamily Parvovirinae Genus Bocaparvovirus
Species Bocaparvovirus primate1 Sequence Length 4953
Definition Human bocavirus isolate 22730/05 NS1, NP-1, VP1, and VP2 genes, complete cds. Submission Date 2008-12-17
Strain Name 22730/05 Sampling Date 2005-09
Sampling Country Germany Location Wuerzburg,ChildrensHospitalUniversity, Germany
Submitted By Ewald,M., Neske,F. and Weissbrich,B. Submitting Institution Virology, Institut for Virology and Immunology - University of Wuerzburg, Versbacherstr.7, Wuerzburg 97078, Germany
Original Host Homo sapiens Sample Type plasma samples
Standardized Host Name Homo sapiens Host Tax Rank species
Standardized Host Class Mammalia Standardized Host Order Primates
Environmental Origin - N/A - Reagent Origin - N/A -
Number Reagent/Consumables Purpose PMID/Url Reference
1 Baculovirus vectors Cloning and expressing capsid proteins VP1 of WUPyV and KIPyV and VP2 of HBoV in Sf9 insect cells. 20646318 View Related Publication
2 Sf9 insect cells Host cells for the expression of recombinant baculoviruses.
3 Pfu Polymerase (Fermentas, St. Leon Rot, Germany) Amplification of the VP2 gene of HBoV and VP1 genes of WUPyV and KIPyV.
4 DNA primers BoV3446s and BoV5113a, WU1673s/WU2782a, KI1501s/KI2727a Specific primers used to amplify the VP2 gene of HBoV, and VP1 genes of WUPyV and KIPyV respectively.
5 TOPO cloning kit (Invitrogen) Insertion of PCR amplicons into the pENTR vector.
6 Chemically competent E.coli TOP10 cells (Invitrogen) Transformation of plasmid containing VP genes.
7 QIAprepSpin Miniprep Kit (Qiagen, Hilden, Germany) Extraction of plasmids from overnight cultures.
8 Cellfectin reagent (Invitrogen) Transfection of recombinant baculovirus DNA into Sf9 cells.
9 TC-100 growth medium (Lonza, Basel, Switzerland) with supplements Cultivation of transfected Sf9 cells.
10 Fetal calf serum, penicillin, streptomycin, amphotericin B, and ganciclovir Supplements added to the TC-100 growth medium for optimal cell growth and selection.
11 Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) Separation of expressed proteins for confirmation.
12 HRP-conjugated mouse-anti-V5-antibody (Invitrogen) Immunoblot detection of expressed proteins.
13 SuperSignal West Pico chemiluminescence substrate (Pierce, Rockford, USA) Visualization of immunoblot bands.
14 Fluoresceine-conjugated goat-anti-human-IgG (Invitrogen) Detection of bound human IgG antibodies in immunofluorescence assay (IFA).
15 Evans blue (Mast Diagnostik, Reinfeld, Germany) Counterstaining in IFA.
16 Phosphate buffered saline (PBS) Washing Sf9 cells and diluting samples.
17 Cold acetone Fixation of Sf9 cells on glass slides.
18 Cell lysates Absorption of plasma samples for specificity tests.
19 GraphPad Prism (version 3.0c for Mac, GraphPad Software, San Diego, USA), SPSS (version 16 for windows, SPSS, Chicago, USA) Statistical analysis of the data.