title: Toolkit for visualization of the cellular structure and organelles in Aspergillus niger.
abstract: Aspergillus niger is a filamentous fungus that is extensively used in industrial fermentations for protein expression and the production of organic acids. Inherent biosynthetic capabilities, such as the capacity to secrete these biomolecules in high amounts, make A. niger an attractive production host. Although A. niger is renowned for this ability, the knowledge of the molecular components that underlie its production capacity, intercellular trafficking processes and secretion mechanisms is far from complete. Here, we introduce a standardized set of tools, consisting of an N-terminal GFP-actin fusion and codon optimized eforRed chromoprotein. Expression of the GFP-actin construct facilitates visualization of the actin filaments of the cytoskeleton, whereas expression of the chromoprotein construct results in a clearly distinguishable red phenotype. These experimentally validated constructs constitute the first set of standardized A. niger biomarkers, which can be used to study morphology, intercellular trafficking, and secretion phenomena.
authors.affiliation: Wageningen_UR iGEM team 2013, ‡Laboratory of Systems and Synthetic Biology, §Laboratory of Microbiology, Wageningen University , 6708 PB Wageningen, The Netherlands.
authors.forename: Emiel B J Ten
authors.initials: EB
authors.lastname: Buren
authors.forename: Michiel A P
authors.initials: MA
authors.lastname: Karrenbelt
authors.forename: Marit
authors.initials: M
authors.lastname: Lingemann
authors.forename: Shreyans
authors.initials: S
authors.lastname: Chordia
authors.forename: Ying
authors.initials: Y
authors.lastname: Deng
authors.forename: JingJing
authors.initials: J
authors.lastname: Hu
authors.forename: Johanna M
authors.initials: JM
authors.lastname: Verest
authors.forename: Vincen
authors.initials: V
authors.lastname: Wu
authors.forename: Teresita J Bello
authors.initials: TJ
authors.lastname: Gonzalez
authors.forename: Ruben G A van
authors.initials: RG
authors.lastname: Heck
authors.forename: Dorett I
authors.initials: DI
authors.lastname: Odoni
authors.forename: Tom
authors.initials: T
authors.lastname: Schonewille
authors.forename: Laura van der
authors.initials: Lv
authors.lastname: Straat
authors.forename: Leo H de
authors.initials: LH
authors.lastname: Graaff
authors.forename: Mark W J van
authors.initials: MW
authors.lastname: Passel
mesh_terms: D000199:Actins / Q000737:chemistry / Q000378:metabolism; D001234:Aspergillus niger / Q000737:chemistry / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D015415:Biomarkers / Q000737:chemistry / Q000378:metabolism; D022082:Cellular Structures / Q000648:ultrastructure*; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000378:metabolism; D015388:Organelles / Q000648:ultrastructure*; D059010:Single-Cell Analysis
chemical_list: D000199:Actins; D015415:Biomarkers; D049452:Green Fluorescent Proteins