Simplified screening for the detection of soluble fusion constructs expressed in <Emphasis Type="Italic">E. coli</Emphasis> using a modular set of vectors |
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Authors: | Annett?Dümmler Ann-Marie?Lawrence Email author" target="_blank">Ario?de MarcoEmail author |
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Institution: | (1) Protein Expression and Purification Facility, EMBL, Meyerhofstr. 1, D-69117 Heidelberg, Germany |
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Abstract: | Background The solubility of recombinant proteins expressed in bacteria is often disappointingly low. Several strategies have been developed
to improve the yield and one of the most common strategies is the fusion of the target protein with a suitable partner. Despite
several reports on the successful use of each of these carriers to increase the solubility of some recombinant proteins, none
of them was always successful and a combinatorial approach seems more efficient to identify the optimal combination for a
specific protein. Therefore, the efficiency of an expression system critically depends on the speed in the identification
of the optimal combination for the suitable fusion candidate in a screening process. This paper describes a set of expression
vectors (pETM) designed for rapid subcloning, expression and subsequent purification using immobilized metal affinity chromatography
(IMAC). |
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Keywords: | |
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