Literature DB >> 8527943

Rapid purification of recombinant green fluorescent protein using the hydrophobic properties of an HPLC size-exclusion column.

J R Deschamps1, C E Miller, K B Ward.   

Abstract

The green fluorescent protein (GFP) of the jelly fish Aequoria victoria was cloned into an Escherichia coli cell line that is a methionine auxotroph. The recombinant GFP (rGFP) was isolated from the cells and purified using a simple procedure consisting of only two chromatographic steps: size-exclusion chromatography and ion-exchange HPLC. Due to the hydrophobic nature of the protein, the surface characteristics of the HPLC size column, and the high initial salt concentration, the rGFP sticks to the size column and is eluted by reducing the salt concentration. Due to this unique behavior the purification procedure can readily be scaled to handle larger quantities of rGFP.

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Year:  1995        PMID： 8527943     DOI： 10.1006/prep.1995.1073

Source DB:  PubMed          Journal:  Protein Expr Purif        ISSN： 1046-5928            Impact factor:   1.650


  8 in total

1.  Mechanism and energetics of green fluorescent protein chromophore synthesis revealed by trapped intermediate structures.

Authors:  David P Barondeau; Christopher D Putnam; Carey J Kassmann; John A Tainer; Elizabeth D Getzoff
Journal:  Proc Natl Acad Sci U S A       Date:  2003-10-01       Impact factor: 11.205

2.  Analysis of Fluorescent Proteins with a Nanoparticle Probe.

Authors:  Francisco A Fernandez-Lima; Michael J Eller; J Daniel Debord; Michaella J Levy; Stanislav V Verkhoturov; Serge Della-Negra; Emile A Schweikert
Journal:  J Phys Chem Lett       Date:  2012-02-02       Impact factor: 6.475

Review 3.  Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.

Authors:  H M Davey; D B Kell
Journal:  Microbiol Rev       Date:  1996-12

4.  Surface display of a single-domain antibody library on Gram-positive bacteria.

Authors:  Filippa Fleetwood; Nick Devoogdt; Mireille Pellis; Ulrich Wernery; Serge Muyldermans; Stefan Ståhl; John Löfblom
Journal:  Cell Mol Life Sci       Date:  2012-10-13       Impact factor: 9.261

5.  Low-mutation-rate, reduced-genome Escherichia coli: an improved host for faithful maintenance of engineered genetic constructs.

Authors:  Bálint Csörgo; Tamás Fehér; Edit Tímár; Frederick R Blattner; György Pósfai
Journal:  Microb Cell Fact       Date:  2012-01-20       Impact factor: 5.328

6.  Semi-automated hydrophobic interaction chromatography column scouting used in the two-step purification of recombinant green fluorescent protein.

Authors:  Orrin J Stone; Kelly M Biette; Patrick J M Murphy
Journal:  PLoS One       Date:  2014-09-25       Impact factor: 3.240

7.  Purification of the Recombinant Green Fluorescent Protein Using Aqueous Two-Phase System Composed of Recyclable CO2-Based Alkyl Carbamate Ionic Liquid.

Authors:  Cher Pin Song; Poh En Liew; Zora Teh; Schian Pei Lim; Pau Loke Show; Chien Wei Ooi
Journal:  Front Chem       Date:  2018-10-31       Impact factor: 5.221

8.  Improved GFP Variants to Study Gene Expression in Haloarchaea.

Authors:  Johannes Born; Felicitas Pfeifer
Journal:  Front Microbiol       Date:  2019-05-29       Impact factor: 5.640
  8 in total

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