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Record W1234396245 · doi:10.1385/0-89603-178-0:237

Application of the Firefly Luciferase Reporter Gene

2003· article· en· W1234396245 on OpenAlexaff
Vincent Giguère

Bibliographic record

VenueHumana Press eBooks · 2003
Typearticle
Languageen
FieldBiochemistry, Genetics and Molecular Biology
Topicbioluminescence and chemiluminescence research
Canadian institutionsSickKids FoundationHospital for Sick Children
Fundersnot available
KeywordsReporter geneBiologyPromoterTransactivationGeneMolecular biologyDNA-binding domainBioreporterLuciferaseResponse elementFusion geneTherapeutic gene modulationGene expressionTranscription factorTransfectionGeneticsRegulator gene

Abstract

fetched live from OpenAlex

Reporter genes have become powerful tools to study regulation of gene expression in eukaryotes. In particular, chimeric transcription units generated by the fusion of the appropriate DNA regulatory sequences to reporter genes have led to the identification of a great number of DNA control elements that constitute eukaryotic promoters. Recently, transcription factors that interact with specific DNA control elements have been purified and the cDNAs encoding these DNA-binding proteins have been cloned. Among these factors, the steroid hormone receptors have been shown to belong to a superfamily of transcription factors that regulate gene expression in a liganddependent fashion (1). The cloning of their respective cDNAs provided the opportunity to identifjr the functional domains for hormone binding, DNA binding, and transactivation present in these proteins. To perform these studies, we developed a screening assay that uses cultured cells transfected with two expression vectors (2). The first vector directs the expression of the wild-type or in vitro mutagenized receptor, and the second contains a reporter gene linked to a hormone-responsive promoter. Application of the hormone or a related synthetic drug activates the reporter gene, and the effects of the mutations on the ability of hormone/receptor complex to acti vate gene expression can be assessed. Because of the large number of experiments to be performed in these studies, the use of chloramphenicol acetyl transferase (CAT) (originally used in our cotransfection assay) as a reporter gene proved to be both laborious and onerous. In contrast, the luciferase gene possesses intrinsic qualities that most other reporter genes lack greater sensitivity, ease of use, instantaneous quantification, " environment friendliness" (no radiolabeled compound or organic solvent is used in this assay), and cost efficiency. Here, the properties of the luciferase system are briefly discussed and a simple and rapid protocol used in my laboratory to assay luciferase activity in extract obtained from cultured mammalian cells transfected with a luciferase reporter gene.

Fetched live from OpenAlex and de-inverted. Abstracts are not stored in this database: the inverted indexes are 8.6 GB of the frame’s 9.3 GB of text, and the host has 13 GB free.

How this classification was reachedexpand

Full frame distilled prediction

Teacher imitation

Not calibrated prevalence, not ground truth. Human validation pending. Learned from the 10,348 direct Codex labels and 10,348 direct Gemma labels. Candidate is the union of thresholded teacher heads; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels or direct frontier model labels.

metaresearch head score (Codex)0.000
metaresearch head score (Gemma)0.000
Version: codex-gemma-dda1882f352aValidation status: machine_predicted_unvalidated
Candidate categoriesnone
Consensus categoriesnone
DomainCandidate signal: none · Consensus signal: none
Study designCandidate signal: Bench or experimental · Consensus signal: Bench or experimental
GenreCandidate signal: Empirical · Consensus signal: Empirical
Teacher disagreement score0.087
Threshold uncertainty score0.272

Codex and Gemma teacher scores by category

CategoryCodexGemma
Metaresearch0.0000.000
Meta-epidemiology (narrow)0.0000.000
Meta-epidemiology (broad)0.0000.000
Bibliometrics0.0000.000
Science and technology studies0.0000.000
Scholarly communication0.0000.000
Open science0.0000.000
Research integrity0.0000.000
Insufficient payload (model declined to judge)0.0000.000

Machine scores (provisional)

The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.

Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.

Opus teacher head0.027
GPT teacher head0.279
Teacher spread0.252 · how far apart the two teachers sit on this one work
Validation statusscore_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from it

Classification

machine, unvalidated

Machine predicted; a candidate call from one teacher head, not a consensus.

The models applied no category: nothing in the taxonomy fit this work.
Study designBench or experimental
Domainnot available
GenreEmpirical

How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".

Quick stats

Citations8
Published2003
Admission routes1
Has abstractyes

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