On-chip Escherichia coli culture, purification, and detection of expressed proteins.
Eur Biophys J
; 35(8): 655-62, 2006 Oct.
Article
in En
| MEDLINE
| ID: mdl-16724194
ABSTRACT
In a recent study, we reported the results of a rapid high-throughput expression analysis of the affinity-tagged proteins present in total cell lysates, using a surface plasmon resonance (SPR) imaging protein chip system. In this paper, we describe a novel method, which is able to sequentially carry out a recombinant Escherichia coli culture, as well as the detection and purification of the expressed proteins on a single microwell chip, fabricated on a two-dimensional thin gold film. Following the induction of the protein on the microwell chip, the E. coli cells were lysed on the chip via the addition of lysozymes, and the expressed glutathione S-transferase-fused green fluorescent protein (GST-GFP) was then purified on the chip via affinity interaction with the glutathionylated gold surface of the chip. Finally, the expressed protein was directly detected using the surface plasmon resonance (SPR) imaging system. This system saves a substantial amount of time, experimental resources, and labor, by allowing for the complicated and labor-intensive procedures inherent to the production of recombinant proteins to be conducted on a single microwell chip, simply and economically.
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Collection:
01-internacional
Health context:
3_ND
Database:
MEDLINE
Main subject:
Recombinant Fusion Proteins
/
Escherichia coli
/
Glutathione Transferase
Type of study:
Diagnostic_studies
Language:
En
Journal:
Eur Biophys J
Year:
2006
Document type:
Article