Gateway Protein Expression and Purification

Genes were transferred from pENTR into pDEST-17 using the Gateway recombinase system (Life Technologies). Proteins were expressed and purified as previously described (6 (link)). In short, E. coli Rosetta cells were induced at 30°C with a final concentration of 0.1 mM IPTG for 3 h. His-tagged proteins were purified by affinity chromatography with an ÄKTA Protein Purification System (GE Life Sciences) using a HisTrap TALON crude column (GE Life Science).

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de Lange O., Wolf C., Thiel P., Krüger J., Kleusch C., Kohlbacher O, & Lahaye T. (2015). DNA-binding proteins from marine bacteria expand the known sequence diversity of TALE-like repeats. Nucleic Acids Research, 43(20), 10065-10080.

Publication 2015

ÄKTA Protein Purification System

Affinity chromatography Cells E coli Genes Iptg Protein Recombinase Talon

Corresponding Organization : University of Tübingen

Other organizations : NanoTemper Technologies (Germany)

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Variable analysis

independent variables

Induction of protein expression in E. coli Rosetta cells with 0.1 mM IPTG

dependent variables

Protein expression and purification

control variables

Temperature (30°C)
Induction time (3 hours)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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