{"doi":"10.1002/yea.2931","title":"Construction of a cytosolic firefly luciferase reporter cassette for use in PCR‐mediated gene deletion and fusion in <i>Saccharomyces cerevisiae</i>","abstract":"<jats:title>Abstract</jats:title><jats:p>Monitoring promoter response to environmental changes using reporter systems has provided invaluable information regarding cellular state. With the development of <jats:italic>in vivo</jats:italic> luciferase reporter systems, inexpensive, sensitive and accurate promoter assays have been developed without the variability reported between <jats:italic>in vitro</jats:italic> samplings. Current luciferase reporter systems, however, are largely inflexible to modifications to the promoter of interest. To overcome problems in flexibility and stability of these expression vectors, we report the creation of a novel vector system which introduces a cytosol‐localized <jats:italic>Photinus pyralis</jats:italic> luciferase [<jats:italic>LUC*(−SKL)</jats:italic>] capable of one‐step, <jats:italic>in vivo</jats:italic> measurements into a promoter–reporter system via PCR‐based gene deletion and fusion. After introduction of the reporter under <jats:italic>HUG1</jats:italic> promoter control, cytosolic localization was confirmed by fluorescence microscopy. The dose–response of this novel construct was then compared with that of a similar <jats:italic>HUG1Δ::yEGFP1</jats:italic> promoter–reporter system and shown to give a similar response pattern. Copyright © 2012 John Wiley &amp; Sons, Ltd.</jats:p>","journal":"Yeast","year":2012,"id":20955,"datarank":0.28162961112402757,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"self_citation_contribution":0.24141568686511508,"citation_network_contribution":0.04021392425891247,"self_endowment_contribution":0.24141568686511508,"citer_contribution":0.04021392425891247,"corpus_percentile":null,"corpus_rank":null,"citation_count":4,"citer_count":3,"citers_with_citation_signal":3,"citers_with_endowment":3,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":null,"is_data_producer":false,"deposit_databanks":null,"is_oa":false,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":null,"fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":136736,"name":"C. M. Rome","orcid":null,"position":1,"is_corresponding":false},{"id":136738,"name":"M. A. Hjortsø","orcid":null,"position":2,"is_corresponding":false},{"id":136741,"name":"M. G. Benton","orcid":null,"position":3,"is_corresponding":false},{"id":136735,"name":"W. B. Ainsworth","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"base_score":1.6094379124341003,"endowment":1.6094379124341003,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"23172625","pmcid":null,"openalex_id":"https://openalex.org/W1960333619","authors":[],"funders":[],"total_grants":0,"fwci":0.2687,"citation_percentile":0.5653167,"influential_citations":0,"citation_trend":[{"year":2013,"count":1},{"year":2015,"count":1},{"year":2022,"count":1},{"year":2024,"count":1}],"oa_status":"closed","license":"http://onlinelibrary.wiley.com/termsAndConditions#vor","oa_locations":[{"url":"https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1002%2Fyea.2931","host_type":"publisher"},{"url":"https://onlinelibrary.wiley.com/doi/pdf/10.1002/yea.2931","host_type":"publisher"},{"url":"https://doi.org/10.1002/yea.2931","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/23172625","host_type":"repository"},{"url":"https://repository.lsu.edu/chem_engineering_pubs/273","host_type":"journal"}],"fields_of_study":["bioluminescence and chemiluminescence research","Neurobiology and Insect Physiology Research","Viral Infectious Diseases and Gene Expression in Insects","Biology","Medicine","Engineering","Animals","Cytosol","Enzyme Induction","Fireflies","Gene Deletion","Gene Fusion","Gene Knockout Techniques","Genes, Reporter","Genetic Engineering","Genetic Vectors","Green Fluorescent Proteins","Luciferases, Firefly","Luminescent Agents","Luminescent Measurements","Peptide Elongation Factor 1","Plasmids","Polymerase Chain Reaction","Promoter Regions, Genetic","Recombinant Fusion Proteins","Saccharomyces cerevisiae","Saccharomyces cerevisiae Proteins"],"mesh_terms":["Animals","Cytosol","Enzyme Induction","Genetic Engineering","Genetic Vectors","Luminescent Measurements","Plasmids","Promoter Regions, Genetic","Recombinant Fusion Proteins","Saccharomyces cerevisiae","Polymerase Chain Reaction","Gene Deletion","Genes, Reporter","Peptide Elongation Factor 1","Saccharomyces cerevisiae Proteins","Fireflies","Luminescent Agents","Luciferases, Firefly","Green Fluorescent Proteins","Gene Fusion","Gene Knockout Techniques"],"keywords":["Luciferase","Reporter gene","Biology","Bioreporter","Molecular biology","Promoter","Green fluorescent protein","Computational biology","Cell biology","Gene","Gene expression","Transfection","Biochemistry"],"sdg_mappings":[{"sdg_number":0,"sdg_label":"Life in Land"}],"linked_datasets":[],"clinical_trials":[],"software_tools":[],"database_accessions":[],"source":"live","citation_network_status":"fetched"},"created_at":"2026-06-06T13:48:37.207393Z","pmid":null,"pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}