title: Development of an expression plasmid and its use in genetic manipulation of Lingzhi or Reishi medicinal mushroom, Ganoderma lucidum (higher Basidiomycetes).
abstract: We report the construction of a plasmid, pJW-EXP, designed for the expression of homologous and heterologous genes in Ganoderma lucidum. pJW-EXP was generated from the plasmid pMD19-T by inserting the G. lucidum glyceraldehyde-3-phosphate dehydrogenase gene promoter, the G. lucidum iron-sulfur protein subunit of succinate dehydrogenase gene terminator and the homologous carboxin-resistance gene as selection marker. This expression plasmid can be efficiently transformed into Ganoderma through polyethylene glycol-mediated protoplast transformation. Southern blot analysis showed that most of the integrated DNA appeared as multiple copies in the genome. The applicability of the constructed plasmid was tested by expression of the truncated G. lucidum 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMGR) gene that encodes the catalytic domain of HMGR. Overexpression of the truncated HMGR gene, which is a key gene in the biosynthetic pathway of the antitumor compounds, ganoderic acids, increased the transcription of the HMGR gene and enhanced ganoderic acid accumulation. pJW-EXP can serve as a useful tool in the genetic improvement and metabolic engineering of Ganoderma.
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Xuya
authors.initials: X
authors.lastname: Yu
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Sen-Lin
authors.initials: SL
authors.lastname: Ji
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Yi-Long
authors.initials: YL
authors.lastname: He
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Meng-Fei
authors.initials: MF
authors.lastname: Ren
authors.affiliation: Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming, 650500, China.
authors.forename: Jun-Wei
authors.initials: JW
authors.lastname: Xu
mesh_terms: D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D018628:Gene Dosage; D015870:Gene Expression; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D020940:Reishi / Q000235:genetics* / Q000378:metabolism; D014170:Transformation, Genetic
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins