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Title 

On-chip Escherichia coli culture, purification, and detection of expressed proteins

Authors 

Moonil KimSo Young LeeHyun-Ju ChoiYong Beom ShinSun Ok JungMin-Gon KimBong Hyun Chung

Publisher 

Springer Verlag (Germany)

Issue Date 

2006

Citation 

European Biophysics Journal With Biophysics Letters, vol. 35, no. 8, pp. 655-662

Keywords 

glutathione S-transferase-fused green fluorescent protein (GST-GFP)microwell chipon-chip purificationsurface plasmon resonance (SPR) imaging systemglutathione transferasegreen fluorescent proteinproteinrecombinant proteinbacterium cultureescherichia coli

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.

ISSN 

0175-7571

Link 

http://dx.doi.org/10.1007/s00249-006-0072-8

Appears in Collections

1. Journal Articles > Journal Articles

Registered Date

2019-05-02


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