{"doi":"10.1006/fgbi.2001.1280","title":"Functional Expression and Cellular Localization of a Green Fluorescent Protein-Tagged Proline Transporter in Aspergillus nidulans","abstract":null,"journal":"Fungal Genetics and Biology","year":2001,"id":47517,"datarank":1.1574945551451654,"base_score":3.4339872044851463,"endowment":3.4339872044851463,"self_citation_contribution":0.515098080672772,"citation_network_contribution":0.6423964744723935,"self_endowment_contribution":0.515098080672772,"citer_contribution":0.6423964744723935,"corpus_percentile":null,"corpus_rank":null,"citation_count":30,"citer_count":14,"citers_with_citation_signal":14,"citers_with_endowment":14,"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":219874,"name":"Claudio Scazzocchio","orcid":null,"position":1,"is_corresponding":false},{"id":219875,"name":"Vicky Sophianopoulou","orcid":null,"position":2,"is_corresponding":false},{"id":219873,"name":"Stefanos Tavoularis","orcid":null,"position":0,"is_corresponding":false}],"reference_count":0,"raw_metadata":{"has_enrichment":true,"resolved":true,"title":"Functional Expression and Cellular Localization of a Green Fluorescent Protein-Tagged Proline Transporter in Aspergillus nidulans","abstract":"The PrnB protein is a highly specific proline transporter that belongs to an amino acid transporter family conserved in both prokaryotes and eukaryotes. In this work, we detected and analyzed the cellular localization of PrnB in vivo by means of green fluorescent protein (GFP) fusions. Several prnB-gfp gene fusions, driven by prnB native promoter sequences, were constructed and targeted to the genomic locus of a prnB null mutant. Chimeric proteins containing GFP fused to the C terminus of PrnB through a linker of two, four, or eight amino acids, with low potential to form secondary structure elements, were shown to be functional in vivo. A two-linker fusion results in partial complementation at both 25 and 37 degrees C. A four-linker fusion affords almost full complementation at 25 degrees C but partial complementation at 37 degrees C, whereas the eight-linker fusion results in partial complementation at both temperatures but in no GFP fluorescence. These results show that the number of linker amino acids is critical for the correct expression and/or translocation of PrnB-GFP fused proteins to the plasma membrane and for the fluorescence of the GFP. The expression of the four-linker PrnB-GFP transporter was detected and analyzed in vivo by both conventional fluorescence and confocal laser microscopy. This chimeric protein is localized in the plasma membrane, secondarily in large vacuoles found in the swollen conidial end of the germlings, and in other small intracellular structures observed as fluorescent granules. A strong correlation between known patterns of PrnB expression and intensity of PrnB-GFP fluorescence was observed. This work also demonstrates that the GFP fusion technology is a unique tool with which to study the expression and cellular localization of low-abundance transmembrane transporters expressed from their native promoters.","is_dataset_classified":null,"base_score":3.4339872044851463,"endowment":3.4339872044851463,"datacite_reuse_total":0,"file_count":0,"downloads":0,"views":0,"has_version_chain":false,"is_dataset":false,"is_oa":false,"pmid":"11456464","pmcid":null,"openalex_id":"https://openalex.org/W2079909060","authors":[],"funders":[],"total_grants":0,"fwci":0.5796,"citation_percentile":0.65561711,"influential_citations":2,"citation_trend":[{"year":2012,"count":1},{"year":2013,"count":2},{"year":2015,"count":1},{"year":2016,"count":1},{"year":2017,"count":1},{"year":2018,"count":1},{"year":2019,"count":1},{"year":2026,"count":1}],"oa_status":"closed","license":"https://www.elsevier.com/tdm/userlicense/1.0/","oa_locations":[{"url":"https://api.elsevier.com/content/article/PII:S1087184501912800?httpAccept=text/xml","host_type":"publisher"},{"url":"https://api.elsevier.com/content/article/PII:S1087184501912800?httpAccept=text/plain","host_type":"publisher"},{"url":"https://doi.org/10.1006/fgbi.2001.1280","host_type":"journal"},{"url":"https://pubmed.ncbi.nlm.nih.gov/11456464","host_type":"repository"}],"fields_of_study":["Fungal and yeast genetics research","Microbial Natural Products and Biosynthesis","Bacterial Genetics and Biotechnology","Biology","Medicine","Amino Acid Transport Systems, Neutral","Aspergillus nidulans","Biological Transport","Carrier Proteins","Cell Membrane","Fluorometry","Fungal Proteins","Gene Expression Regulation, Fungal","Green Fluorescent Proteins","Hot Temperature","Indicators and Reagents","Kinetics","Luminescent Proteins","Plasmids","Recombinant Fusion Proteins"],"mesh_terms":["Aspergillus nidulans","Biological Transport","Carrier Proteins","Cell Membrane","Fluorometry","Fungal Proteins","Hot Temperature","Indicators and Reagents","Kinetics","Luminescent Proteins","Plasmids","Recombinant Fusion Proteins","Gene Expression Regulation, Fungal","Amino Acid Transport Systems, Neutral","Green Fluorescent Proteins"],"keywords":["Green fluorescent protein","Complementation","Biology","Bimolecular fluorescence complementation","Fusion protein","Linker","Biochemistry","Amino acid","Amino acid transporter","Aequorea victoria","Yellow fluorescent protein","Transport protein","Protein-fragment complementation assay","Molecular biology","Cell biology","Mutant","Transporter","Gene","Recombinant DNA"],"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":"2026-07-18T19:05:55.174433Z","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":[]}