title: Cyclin B1 destruction box-mediated protein instability: the enhanced sensitivity of fluorescent-protein-based reporter gene system.
abstract: The periodic expression and destruction of several cyclins are the most important steps for the exact regulation of cell cycle. Cyclins are degraded by the ubiquitin-proteasome system during cell cycle. Besides, a short sequence near the N-terminal of cyclin B called the destruction box (D-box; CDB) is also required. Fluorescent-protein-based reporter gene system is insensitive to analysis because of the overly stable fluorescent proteins. Therefore, in this study, we use human CDB fused with both enhanced green fluorescent protein (EGFP) at C-terminus and red fluorescent protein (RFP, DsRed) at N-terminus in the transfected human melanoma cells to examine the effects of CDB on different fluorescent proteins. Our results indicated that CDB-fused fluorescent protein can be used to examine the slight gene regulations in the reporter gene system and have the potential to be the system for screening of functional compounds in the future.
authors.affiliation: Department of Cosmetic Science, Providence University, No. 200, Section 7, Taiwan Boulevard, Shalu District, Taichung 43301, Taiwan.
authors.forename: Chao-Hsun
authors.initials: CH
authors.lastname: Yang
authors.affiliation: Department of Cosmetic Science, Providence University, No. 200, Section 7, Taiwan Boulevard, Shalu District, Taichung 43301, Taiwan.
authors.forename: Wan-Ting
authors.initials: WT
authors.lastname: Kuo
authors.affiliation: Department of Cosmetic Science, Providence University, No. 200, Section 7, Taiwan Boulevard, Shalu District, Taichung 43301, Taiwan.
authors.forename: Yun-Ting
authors.initials: YT
authors.lastname: Chuang
authors.affiliation: Department of Cosmetic Science, Providence University, No. 200, Section 7, Taiwan Boulevard, Shalu District, Taichung 43301, Taiwan ; Graduate Institute of Biotechnology, National Chung Hsing University, 250 Kuo-Kuang Road, Taichung 40227, Taiwan.
authors.forename: Cheng-Yu
authors.initials: CY
authors.lastname: Chen
authors.affiliation: Department of Cosmetic Science, Providence University, No. 200, Section 7, Taiwan Boulevard, Shalu District, Taichung 43301, Taiwan.
authors.forename: Chih-Chien
authors.initials: CC
authors.lastname: Lin
mesh_terms: D000595:Amino Acid Sequence / Q000235:genetics*; D002453:Cell Cycle / Q000235:genetics*; D045744:Cell Line, Tumor; D056744:Cyclin B1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D017930:Genes, Reporter / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics; D006801:Humans; D008164:Luminescent Proteins / Q000737:chemistry / Q000235:genetics; D055550:Protein Stability; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D017384:Sequence Deletion; D000097573:Red Fluorescent Protein
chemical_list: D056744:Cyclin B1; D049452:Green Fluorescent Proteins; D008164:Luminescent Proteins; D011994:Recombinant Proteins; D000097573:Red Fluorescent Protein