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231224s2015 xx |||||o 00| ||eng c |
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7 |
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|a 10.2144/000114265
|2 doi
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|a eng
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| 100 |
1 |
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|a Coren, Lori V
|e verfasserin
|4 aut
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| 245 |
1 |
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|a Production of retroviral constructs for effective transfer and expression of T-cell receptor genes using Golden Gate cloning
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| 264 |
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|c 2015
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| 336 |
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|a Text
|b txt
|2 rdacontent
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|a ƒaComputermedien
|b c
|2 rdamedia
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|a ƒa Online-Ressource
|b cr
|2 rdacarrier
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| 500 |
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|a Date Completed 30.11.2015
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|a Date Revised 15.06.2024
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|a published: Electronic-eCollection
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|a Citation Status MEDLINE
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|a Here we present an improved strategy for producing T-cell receptor (TCR)-expressing retroviral vectors using a Golden Gate cloning strategy. This method takes advantage of the modular nature of TCR genes by directly amplifying TCR α and β variable regions from RNA or cDNA, then cloning and fusing them with their respective constant region genes resident in a retroviral TCR expression vector. Our one-step approach greatly streamlines the TCR vector production process in comparison to the traditional three-step procedure that typically involves cloning whole TCR genes, producing a TCR expression cassette, and constructing a retroviral construct. To date, we have generated TCR vectors that transferred seven functional human/rhesus macaque TCRs into primary T cells. The approach also holds promise for the assembly of other genes with defined variable regions, such as immunoglobulins
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4 |
|a Journal Article
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4 |
|a Research Support, N.I.H., Extramural
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| 650 |
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4 |
|a Golden Gate cloning
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| 650 |
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4 |
|a T-cell receptor
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| 650 |
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4 |
|a gene engineering
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| 650 |
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4 |
|a retroviral vector
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| 650 |
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7 |
|a Receptors, Antigen, T-Cell, alpha-beta
|2 NLM
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| 700 |
1 |
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|a Jain, Sumiti
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Trivett, Matthew T
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Ohlen, Claes
|e verfasserin
|4 aut
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| 700 |
1 |
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|a Ott, David E
|e verfasserin
|4 aut
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| 773 |
0 |
8 |
|i Enthalten in
|t BioTechniques
|d 1988
|g 58(2015), 3 vom: 15. März, Seite 135-9
|w (DE-627)NLM012627046
|x 1940-9818
|7 nnns
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| 773 |
1 |
8 |
|g volume:58
|g year:2015
|g number:3
|g day:15
|g month:03
|g pages:135-9
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| 856 |
4 |
0 |
|u http://dx.doi.org/10.2144/000114265
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