title: Maize Activator transposase expressed in Saccharomyces cerevisiae from a genomic clone: detection via Elisa, and proposed use in complementation studies.
abstract: The maize Activator (Ac) transposase (TPase) was expressed as a histidine (His)-tagged protein in Saccharomyces cerevisiae from a full-length genomic clone. Expression was demonstrated via the highly specific nickel-coated Elisa plate method, using an anti-His antibody and 2 separate anti-Ac TPase antibodies, to Ac residues 103-465 and 189-807. Ac TPase expression in S. cerevisiae is important for two reasons: (a) because the expression from a genomic clone herein permits the future study of RNA splicing mechanisms in common between maize and yeast systems, and (b) because a yeast system can easily be used for demonstrating complementation of function. Thus, such transformed yeast systems could be used in future to experimentally test whether Ac TPase could complement various yeast mutations. Specifically, Ac TPase may be able to complement (i.e., provide the same function) to yeast transcription factor mutants, or, to genes mutated in other essential yeast functions. If confirmed, this would lend support to Barbara McClintock's hypothesis that transposable elements can serve as 'controlling elements' within the genome, by their ability to supplement other essential genes' functions, as needed. Work herein is contrasted with existing studies on Ac in yeast.
authors.affiliation: Genomic Regulation, LLC, St. Louis, Mo. 63146, USA. amacrae@swbell.net
authors.forename: Amy F
authors.initials: AF
authors.lastname: MacRae
mesh_terms: D001709:Biotechnology / Q000379:methods; D003001:Cloning, Molecular; D004797:Enzyme-Linked Immunosorbent Assay; D005816:Genetic Complementation Test; D016681:Genome, Fungal; D006639:Histidine / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D014170:Transformation, Genetic; D019895:Transposases / Q000235:genetics / Q000378:metabolism*; D003313:Zea mays / Q000201:enzymology* / Q000235:genetics*
chemical_list: D006639:Histidine; D019895:Transposases