Open AccessStandardization of Functional Reporter and Antibiotic Resistance Cassettes to Facilitate the Genetic Engineering of Filamentous Fungi
Author(s) -
Swati Sureka,
Arun K. Chakravorty,
Eric C. Holmes,
Olga Spassibojko,
Nupur Bhatt,
Dongliang Wu,
B. Gillian Turgeon
Publication year - 2014
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/sb5000143
Subject(s) - biology , aspergillus nidulans , gene , hygromycin b , computational biology , microbiology and biotechnology , genetics , plasmid , mutant
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.