Evidence map›Paper›PMID 22639380›Full record

ArticleIUBMB life2012

Fluorescent protein engineering by in vivo site-directed mutagenesis.

Melvys Valledor, Qinghua Hu, Paul Schiller, Richard S Myers

Open access · greenAbstract read
In one paragraph

Article in IUBMB life, 2012. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 4 papers.

0numbers the graph read from it
0cells of the map it votes in
4citing papers in PubMed
0.3field-weighted citation impact, top 43% of its field
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

Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.

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

4 citing papers in PubMed, 8 citations in OpenAlex.

  1. Deep Learning-Assisted Design of Novel Promoters inAdvanced genetics (Hoboken, N.J.) · 2023
    Article
  2. Herpes simplex virus 1 ICP8 mutant lacking annealing activity is deficient for viral DNA replication.Proceedings of the National Academy of Sciences of the United States of America · 2019
    Article
  3. Article
  4. Review
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

4 authors at 2 institutions in 1 country.

Melvys ValledorDepartment of Biochemistry and Molecular Biology, University of Miami Miller School of Medicine, Miami, FL, USA.
Qinghua Hu
Paul Schiller
Richard S Myers
Geriatric Research Education and Clinical Center · USUniversity of Miami · US

Funding

University of Miami Developmental Center for AIDS Research (D-CFAR)P30AI073961 · NIAID · UNIVERSITY OF MIAMI SCHOOL OF MEDICINE · PI Rana Chakraborty · 2007 to 2026
$33.8M
Human Stem Cell RecombineeringF31GM089125 · NIGMS · UNIVERSITY OF MIAMI SCHOOL OF MEDICINE · PI VALLEDOR CEBALLOS, MELVYS · 2009 to 2012
$151k
NIAID NIH HHS P30 AI073961NIGMS NIH HHS F31 GM089125NIGMS NIH HHS F31-GM089125-01
6 · The paper itself

Abstract

In vivo site-directed mutagenesis by single-stranded deoxyribonucleic acid recombineering is a facile method to change the color of fluorescent proteins (FPs) without cloning. Two different starting alleles of GFP were targeted for mutagenesis: gfpmut3* residing in the Escherichia coli genome and egfp carried by a bacterial/mammalian dual expression lentiviral plasmid vector. Fluorescent protein spectra were shifted by subtle modification of the chromophore region and residues interacting with the chromophore of the FP. Eight different FPs (Violeta, Azure, Aqua, Mar, Celeste, Amarillo, Mostaza, and Bronze) were isolated and shown to be useful in multicolor imaging and flow cytometry of bacteria and transgenic human stem cells. To make in vivo site-directed mutagenesis more efficient, the recombineering method was optimized using the fluorescence change as a sensitive quantitative assay for recombination. A set of rules to simplify mutant isolation by recombineering is provided.

Indexed as

Directed Molecular EvolutionEscherichia coliFlow CytometryFluorescenceGreen Fluorescent ProteinsHEK293 CellsHematopoietic Stem CellsHumansLuminescent ProteinsMaleMicroscopy, ConfocalMutagenesis, Site-DirectedPlasmidsProtein EngineeringSpectrometry, FluorescenceYoung AdultGreen Fluorescent ProteinsLuminescent Proteins

Identifiers

PMID22639380
PMCPMC3415465
OpenAlexW1971032828

What OpenQuestion holds

Textmetadata
LicenceTDM
Read underepoch 390

Registered trials

None linked

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.