title: [Expression of peaT1 gene from Alternaria tenuissima in Pichia pastoris and its function].
abstract: In this study, peaT1 gene was subcloned into the Pichia pastoris expression vector pPIC9K, which contained both the methanol-inducible promoter and the transcription terminator of the AOX1 gene, resulting the plasmid pPIC9K-peaT1. The recombinant plasmid was linearized by Sal I or Bgl II and transformed into P. pastoris GS115 by electroporation method. Recombinant strain was screened by Minimal Dextrose Medium and further confirmed by PCR. The gene was in frame integrated into the Pichia genome through homologous recombination, resulting the recombinant strain. Regulated by the alpha-Factor, promoter of AOX1 gene and termination signal of yeast genomic, the recombinant protein was expressed and secreted into the supernatant. The SDS-PAGE analysis indicated that the apparent molecular weight of target protein was about 35 kD. Bioassay results showed that the inhibition rate of the expressed protein against TMV was 30.37%. The supernatant was collected and then purified by anion exchange chromatography. This protein can promote seedling growth of wheat obviously.
authors.affiliation: State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing 100081, China.
authors.forename: Yanfeng
authors.initials: Y
authors.lastname: Liu
authors.forename: Hongmei
authors.initials: H
authors.lastname: Zeng
authors.forename: Shanjiang
authors.initials: S
authors.lastname: Yu
authors.forename: Xiufen
authors.initials: X
authors.lastname: Yang
authors.forename: Jianjun
authors.initials: J
authors.lastname: Mao
authors.forename: Dewen
authors.initials: D
authors.lastname: Qiu
mesh_terms: D000528:Alternaria / Q000235:genetics*; D005656:Fungal Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology; D010843:Pichia / Q000235:genetics* / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D014908:Triticum / Q000187:drug effects / Q000254:growth & development
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins