Evidence map›Paper›PMID 35773582›Full record

ArticleMethods in molecular biology (Clifton, N.J.)2022

Production and Purification of a GFP-Tagged ABC Transporter CaCdr1p.

Jorgaq Pata, Alexis Moreno, Sandrine Magnard, Atanu Banerjee, Rajendra Prasad, Pierre Falson

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Article in Methods in molecular biology (Clifton, N.J.), 2022. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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0citing papers in PubMed
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1 · What the graph read from it

What it found

Each row is one number read from the abstract, on the scale the paper reported it, with its interval. Left of the dashed line favours the treatment, right favours the comparator. Under each row is the sentence it came from. New to these charts? A ten-minute tutorial.

The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.

2 · The registry

The trial behind it

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Neither the registry nor the abstract names a trial number. If this is a trial report, that itself is worth knowing.

3 · Its place in the literature

Who cites it

0 citing papers in PubMed.

No citing paper in PubMed yet.

4 · The record

Corrections and comments

PubMed lists nothing against this paper. Absence here is not a guarantee, only a check that was made.

5 · Who and what money

Authors and funding

6 authors.

Jorgaq PataDrug Resistance and Membrane Proteins Team, CNRS - Lyon 1 University UMR 5086, Institut de Biologie et Chimie des Protéines, Lyon, France.
Alexis MorenoCalixar, Lyon, France.
Sandrine MagnardDrug Resistance and Membrane Proteins Team, CNRS - Lyon 1 University UMR 5086, Institut de Biologie et Chimie des Protéines, Lyon, France.
Atanu BanerjeeAmity Institute of Biotechnology, Amity University Haryana, Gurgaon, India.
Rajendra PrasadAmity Institute of Biotechnology, Amity University Haryana, Gurgaon, India.
Pierre FalsonDrug Resistance and Membrane Proteins Team, CNRS - Lyon 1 University UMR 5086, Institut de Biologie et Chimie des Protéines, Lyon, France. pierre.falson@univ-lyon1.fr.ORCID 0000-0002-9760-4577

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

Abstract

The production and purification are the first steps required in any functional or structural study of a protein of interest. In the case of membrane proteins, these tasks can be difficult due to low expression levels and the necessity to extract them from their membrane environment. This chapter describes a convenient method based on GFP tagged to the membrane protein to facilitates these steps. Production is carried out in the yeast S. cerevisiae and purification steps are carried out and monitored taking advantage of an anti-GFP nanobody. We show how GFP can be a very helpful tool for controlling the correct addressing of the protein and for probing and optimizing purification. These methods are described here for producing and purifying CaCdr1p, an ABC exporter conferring multiantifungal resistance to C. albicans. This purification method can be amenable to any other GFP-tagged protein.

Indexed as

ATP-Binding Cassette TransportersSaccharomyces cerevisiaeCandida albicansGreen Fluorescent ProteinsMembrane ProteinsATP-Binding Cassette TransportersGreen Fluorescent ProteinsMembrane ProteinsABC exporterAnti-GFP nanobodyCaCdr1pGFP tagMembrane proteinProduction in yeastS. cerevisiae

Identifiers

PMID35773582

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Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.