title: [Agrobacterium tumefaciens mediated transformation of Aspergillus niger conidiospores and sequence analysis of T-DNA insertion site in conidiation-defective mutant].
abstract: OBJECTIVE
  Development of a method of transforming Aspergillus niger conidiospores with Agrobacterium tumefaciens T-DNA system and using it as a tool for genome annotation via construction a T-DNA insertion mutant library of A. niger.
  
  
  METHODS
  A. tumefaciens EHA105 carrying the binary vector pCAMBIA1301 was used to transform A. niger conidiospores under induction conditions. Hygromycin resistant transformants were screened on the selected medium. Sequence analysis was performed using inverse PCR method in stable mutants.
  
  
  RESULTS
  In total, 193 hygromycin resistant stable transformants were obtained and the transformation rate was about 5.6 x 10(2) transformants /10(8) conidiospores. Obvious morphological changes were observed in some mutants and sequence analysis of T-DNA inserted site was performed in one conidiation-defective mutant. The result showed that the interrupted gene belonged to major facilitator superfamily (MFS).
  
  
  CONCLUSIONS
  Our results indicated that the transformation system of A. niger conidiospores set up in this study, combing with sequence analysis of T-DNA insertion site, might be an effective way for genome annotation in A. niger.
authors.affiliation: Key Laboratory of Industrial Microbiology, Ministry of Education, Tianjin Key Laboratory of Industrial Microbiology, College of Biotechnology, Tianjin University of Science & Technology, Tianjin 300457, China. huangwen@mail.tust.cn
authors.forename: Wen
authors.initials: W
authors.lastname: Huang
authors.forename: Hongjiang
authors.initials: H
authors.lastname: Yang
authors.forename: Huibin
authors.initials: H
authors.lastname: Qin
mesh_terms: D016960:Agrobacterium tumefaciens / Q000235:genetics*; D001234:Aspergillus niger / Q000235:genetics* / Q000254:growth & development*; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics*; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D009154:Mutation; D017422:Sequence Analysis, DNA; D013172:Spores, Fungal / Q000235:genetics / Q000254:growth & development; D014170:Transformation, Genetic
chemical_list: D004269:DNA, Bacterial; C036483:T-DNA