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Implementation of s...
Implementation of semi-automated cloning and prokaryotic expression screening : the impact of SPINE
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Alzari, Pedro M. (author)
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Berglund, H. (author)
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Berrow, N. S. (author)
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Blagova, E. (author)
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Busso, D. (author)
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Cambillau, C. (author)
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Campanacci, V. (author)
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Christodoulou, E. (author)
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Eiler, S. (author)
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Fogg, M. J. (author)
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Folkers, G. (author)
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Geerlof, A. (author)
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Hart, D. (author)
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Haouz, A. (author)
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Herman, M. D. (author)
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Macieira, S. (author)
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- Nordlund, Pär (author)
- Karolinska Institutet
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Perrakis, A. (author)
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Quevillon-Cheruel, S. (author)
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Tarandeau, F. (author)
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van Tilbeurgh, H. (author)
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Unger, T. (author)
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Luna-Vargas, M. P. A. (author)
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Velarde, M. (author)
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Willmanns, M. (author)
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Owens, Raymond J. (author)
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(creator_code:org_t)
- 2006
- 2006
- English.
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In: Acta Crystallographica Section D. - 0907-4449 .- 1399-0047. ; 62, s. 1103-1113
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
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- The implementation of high- throughput ( HTP) cloning and expression screening in Escherichia coli by 14 laboratories in the Structural Proteomics In Europe ( SPINE) consortium is described. Cloning efficiencies of greater than 80% have been achieved for the three non- ligation- based cloning techniques used, namely Gateway, ligation- indendent cloning of PCR products ( LIC- PCR) and In- Fusion, with LIC- PCR emerging as the most cost- effective. On average, two constructs have been made for each of the approximately 1700 protein targets selected by SPINE for protein production. Overall, HTP expression screening in E. coli has yielded 32% soluble constructs, with at least one for 70% of the targets. In addition to the implementation of HTP cloning and expression screening, the development of two novel technologies is described, namely library- based screening for soluble constructs and parallel small- scale high- density fermentation.
Subject headings
- NATURVETENSKAP -- Biologi (hsv//swe)
- NATURAL SCIENCES -- Biological Sciences (hsv//eng)
Keyword
- Biology
- Biologi
Publication and Content Type
- ref (subject category)
- art (subject category)
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- By the author/editor
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Alzari, Pedro M.
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Berglund, H.
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Berrow, N. S.
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Blagova, E.
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Busso, D.
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Cambillau, C.
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show more...
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Campanacci, V.
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Christodoulou, E ...
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Eiler, S.
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Fogg, M. J.
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Folkers, G.
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Geerlof, A.
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Hart, D.
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Haouz, A.
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Herman, M. D.
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Macieira, S.
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Nordlund, Pär
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Perrakis, A.
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Quevillon-Cherue ...
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Tarandeau, F.
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van Tilbeurgh, H ...
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Unger, T.
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Luna-Vargas, M. ...
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Velarde, M.
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Willmanns, M.
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Owens, Raymond J ...
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show less...
- About the subject
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- NATURAL SCIENCES
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NATURAL SCIENCES
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and Biological Scien ...
- Articles in the publication
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Acta Crystallogr ...
- By the university
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Royal Institute of Technology
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Karolinska Institutet