title: Induction and construct UV protective yeast plasmid.
abstract: In this study, we apply concepts of synthetic biology in combination with conventional methods to assemble different genetic components to construct yeast resistant to UV radiation, and to induce production of anti-UV proteins. This work combines sequences of different promoters, STRESS-proteins, heat shock protein (HSP), kinase proteins, alcohol dehydrogenase protein (ADH), ribosomal binding sites, fluorescent reporter proteins, terminators, and a synthetic ribosomal switch. The aim of this investigation was to induce an anti-UV proteins, and to construct an anti-UV yeast plasmid to be used for protection of skin cells against UV radiation. This investigation demonstrates induction and construction of anti-UV genes and production of their corresponding proteins. Cultures of Saccharomyces cerevisiae (ATCC # 66348) were exposed to short-wave UV radiation and were then subjected to time-PCR to assess specific gene expression. Proteins were identified using two dimensional difference gel electrophoresis (2D DIGE) and LC-MS/MS. Different up-regulated and down-regulated proteins were identified. Highly expressed identified proteins were cloned into S. cerevisiae using a synthetic biology approach. Extracts from UV-induced genetically transformed yeasts were used to protect skin cell cultures (ATCC #2522-CRL) in vitro. Both microscopic analysis and an apoptosis assay showed protection of the skin cell cultures against UV radiation.
authors.affiliation: Prairie View A&M University, CARC, Member of the Texas A&M University System, Prairie View, TX 77446, USA. olimpa@aol.com
authors.forename: Raul
authors.initials: R
authors.lastname: Cuero
authors.forename: David S
authors.initials: DS
authors.lastname: McKay
mesh_terms: D017209:Apoptosis; D001483:Base Sequence; D002478:Cells, Cultured; D003001:Cloning, Molecular; D015536:Down-Regulation; D015966:Gene Expression Regulation, Fungal; D005800:Genes, Fungal; D008969:Molecular Sequence Data; D010957:Plasmids; D012045:Regulatory Sequences, Nucleic Acid; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism / Q000528:radiation effects*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism; D058615:Synthetic Biology; D053719:Tandem Mass Spectrometry; D014162:Transfection; D014466:Ultraviolet Rays; D015854:Up-Regulation
chemical_list: D029701:Saccharomyces cerevisiae Proteins