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  • Alzari, Pedro M. (author)

Implementation of semi-automated cloning and prokaryotic expression screening : the impact of SPINE

  • Article/chapterEnglish2006

Publisher, publication year, extent ...

  • 2006
  • printrdacarrier

Numbers

  • LIBRIS-ID:oai:DiVA.org:kth-37384
  • https://urn.kb.se/resolve?urn=urn:nbn:se:kth:diva-37384URI
  • https://doi.org/10.1107/S0907444906029775DOI
  • http://kipublications.ki.se/Default.aspx?queryparsed=id:1942817URI

Supplementary language notes

  • Language:English
  • Summary in:English

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Classification

  • Subject category:ref swepub-contenttype
  • Subject category:art swepub-publicationtype

Notes

  • 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 and genre

Added entries (persons, corporate bodies, meetings, titles ...)

  • Berglund, H. (author)
  • Berrow, N. S. (author)
  • Blagova, E. (author)
  • Busso, D. (author)
  • Cambillau, C. (author)
  • Campanacci, V. (author)
  • Christodoulou, E. (author)
  • Eiler, S. (author)
  • Fogg, M. J. (author)
  • Folkers, G. (author)
  • Geerlof, A. (author)
  • Hart, D. (author)
  • Haouz, A. (author)
  • Herman, M. D. (author)
  • Macieira, S. (author)
  • Nordlund, PärKarolinska Institutet (author)
  • Perrakis, A. (author)
  • Quevillon-Cheruel, S. (author)
  • Tarandeau, F. (author)
  • van Tilbeurgh, H. (author)
  • Unger, T. (author)
  • Luna-Vargas, M. P. A. (author)
  • Velarde, M. (author)
  • Willmanns, M. (author)
  • Owens, Raymond J. (author)
  • Karolinska Institutet (creator_code:org_t)

Related titles

  • In:Acta Crystallographica Section D62, s. 1103-11130907-44491399-0047

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