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rdf:type |
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lifeskim:mentions |
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pubmed:issue |
23
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pubmed:dateCreated |
2002-11-4
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pubmed:abstractText |
Production of large quantities of recombinant adeno-associated virus (AAV) is difficult and not cost-effective. To overcome this problem, we have explored the feasibility of creating a recombinant AAV encoding a 6xHis tag on the VP3 capsid protein. We generated a plasmid vector containing a six-His (6xHis)-tagged AAV VP3. A second plasmid vector was generated that contained the full-length AAV capsid capable of producing VP1 and VP2, but not VP3 due to a mutation at position 2809 that encodes the start codon for VP3. These plasmids, necessary for production of AAV, were transfected into 293 cells to generate a 6xHis-tagged VP3mutant recombinant AAV. The 6xHis-tagged VP3 did not affect the formation of AAV virus, and the physical properties of the 6xHis-modified AAV were equivalent to those of wild-type particles. The 6xHis-tagged AAV did not affect the production titer of recombinant AAV and could be used to purify the recombinant AAV using an Ni-nitrilotriacetic acid column. Addition of the 6xHis tag did not alter the viral tropism compared to wild-type AAV. These observations demonstrate the feasibility of producing high-titer AAV containing a 6xHis-tagged AAV VP3 capsid protein and to utilize the 6xHis-tagged VP3 capsid to achieve high-affinity purification of this recombinant AAV.
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pubmed:grant |
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pubmed:commentsCorrections |
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10219758,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10223736,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10470084,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10600510,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10600599,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10708427,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10716335,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10821405,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-10954565,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11044910,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11096446,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11139798,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11199265,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11269337,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11406765,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11407911,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11713191,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11751689,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11767048,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11781658,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-11827486,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-1331503,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-1653762,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-2156265,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-2547998,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-2841488,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-2847413,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-6086948,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-6088786,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-6265925,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-7996133,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-8030266,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-9191943,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-9445046,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-9741431,
http://linkedlifedata.com/resource/pubmed/commentcorrection/12414944-9883842
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pubmed:language |
eng
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pubmed:journal |
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pubmed:citationSubset |
IM
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pubmed:chemical |
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pubmed:status |
MEDLINE
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pubmed:month |
Dec
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pubmed:issn |
0022-538X
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pubmed:author |
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pubmed:issnType |
Print
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pubmed:volume |
76
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
12023-31
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pubmed:dateRevised |
2009-11-18
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pubmed:meshHeading |
pubmed-meshheading:12414944-Base Sequence,
pubmed-meshheading:12414944-Capsid Proteins,
pubmed-meshheading:12414944-Cell Line,
pubmed-meshheading:12414944-DNA, Viral,
pubmed-meshheading:12414944-Dependovirus,
pubmed-meshheading:12414944-Genetic Vectors,
pubmed-meshheading:12414944-HeLa Cells,
pubmed-meshheading:12414944-Humans,
pubmed-meshheading:12414944-Jurkat Cells,
pubmed-meshheading:12414944-Plasmids,
pubmed-meshheading:12414944-Recombinant Proteins,
pubmed-meshheading:12414944-Virus Replication
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pubmed:year |
2002
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pubmed:articleTitle |
Addition of six-His-tagged peptide to the C terminus of adeno-associated virus VP3 does not affect viral tropism or production.
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pubmed:affiliation |
Division of Clinical Immunology and Rheumatology, Department of Medicine, The University of Alabama at Birmingham, Birmingham, Alabama 35294, USA.
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pubmed:publicationType |
Journal Article,
Research Support, U.S. Gov't, P.H.S.,
Research Support, U.S. Gov't, Non-P.H.S.,
Research Support, Non-U.S. Gov't
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