title: Standardization of functional reporter and antibiotic resistance cassettes to facilitate the genetic engineering of filamentous fungi.
abstract: The unique physiological properties of fungi are useful for a myriad of applications, which could greatly benefit from increased control of native pathways and introduction of recombinant genes. However, fungal genetic engineering is still limited in scope and accessibility, largely due to lack of standardization. To help standardize the genetic engineering of filamentous fungi, we created BioBricks of commonly used antibiotic resistance genes, neomycin phosphotransferase (nptII) and hygromycin phosphotransferase (hph), which confer resistance to G418 (Geneticin) and hygromycin B, respectively. Additionally, we created a BioBrick of the constitutive trpC promoter, from the tryptophan biosynthesis pathway of Aspergillus nidulans, and used it to create a composite part including the GFP gene. The functionality of these parts was demonstrated in the model fungal organism Cochliobolus heterostrophus, and as these tools are in modular BioBrick format, they can be easily used to facilitate genetic engineering of other fungal species.
authors.affiliation: Department of Biological and Environmental Engineering, ‡Department of Molecular Biology and Genetics, §Department of Plant Pathology & Plant-Microbe Biology Section, School of Integrative Plant Science, Cornell University , Ithaca, New York 14853, United States.
authors.forename: Swati
authors.initials: S
authors.lastname: Sureka
authors.forename: Arun
authors.initials: A
authors.lastname: Chakravorty
authors.forename: Eric C
authors.initials: EC
authors.lastname: Holmes
authors.forename: Olga
authors.initials: O
authors.lastname: Spassibojko
authors.forename: Nupur
authors.initials: N
authors.lastname: Bhatt
authors.forename: Dongliang
authors.initials: D
authors.lastname: Wu
authors.forename: B Gillian
authors.initials: BG
authors.lastname: Turgeon
mesh_terms: D001203:Ascomycota / Q000235:genetics*; D001233:Aspergillus nidulans / Q000235:genetics; D004352:Drug Resistance, Microbial / Q000235:genetics*; D017930:Genes, Reporter / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods* / Q000592:standards; D010957:Plasmids / Q000235:genetics*; D014162:Transfection