Change search
ReferencesLink to record
Permanent link

Direct link
Implementation of semi-automated cloning and prokaryotic expression screening: the impact of SPINE
Show others and affiliations
2006 (English)In: Acta Crystallographica Section D: Biological Crystallography, ISSN 0907-4449, E-ISSN 1399-0047, Vol. 62, 1103-1113 p.Article in journal (Refereed) Published
Abstract [en]

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.

Place, publisher, year, edition, pages
2006. Vol. 62, 1103-1113 p.
National Category
Biological Sciences
URN: urn:nbn:se:kth:diva-37384DOI: 10.1107/S0907444906029775ISI: 000240618000001PubMedID: 17001088ScopusID: 2-s2.0-33749519973OAI: diva2:433573
Available from: 2011-08-10 Created: 2011-08-10 Last updated: 2012-03-22Bibliographically approved

Open Access in DiVA

No full text

Other links

Publisher's full textPubMedScopus
In the same journal
Acta Crystallographica Section D: Biological Crystallography
Biological Sciences

Search outside of DiVA

GoogleGoogle Scholar
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

Altmetric score

Total: 16 hits
ReferencesLink to record
Permanent link

Direct link