Title
A dedicated vector for efficient library construction and high throughput screening in the hyphal fungus Chrysosporium lucknowense
Author
Verdoes, J.C.
Punt, P.J.
Burlingame, R.
Bartels, J.
van Dijk, R.
Slump, E.
Meens, M.
Joosten, R.
Emalfarb, M.
TNO Kwaliteit van Leven
Publication year
2007
Abstract
A self-replicating vector was designed that enables the construction of complex libraries in the fungus Chiysosporium lucknowense. The circular vector is linearized in vivo and results in a transformation frequency up to 13,000 transformants/ug ofplasmid DNA. Upon prolonged cultivation of the transformants in liquid medium, the linearized vector finally integrates, mainly as a single copy, at the telomere associated region of the chromosomes. Screening for colonies with compact filamentous growth on agar plates resulted in a C. lucknowense strain with significantly reduced viscosity during fermentation. Further medium modification resulted in conditions that enable cultivation, replication, and screening of a large collection of fungal transformants in microtiter plates. This paper describes the successful integration of library construction and screening in an automated setup using different configurations for a hyphal fungal system.
Subject
Biology
Biotechnology
Chrysosporium lucknowense
Fungal high throughput screening
Hyphal fungal system
Libraries
Propagule
Chromosomes
DNA
Fermentation
Fungi
Growth kinetics
Viscosity
Chrysosporium lucknowense
Fungal high throughput screening
Hyphal fungal system
Propagule
Genetic engineering
Chromosomes
DNA
Fermentation
Fungi
Genetic engineering
Growth kinetics
Viscosity
Chrysosporium
Chrysosporium lucknowense
Fungi
To reference this document use:
http://resolver.tudelft.nl/uuid:208a154e-464d-4cb7-bb77-edfd69aa9434
DOI
https://doi.org/10.1089/ind.2007.3.048
TNO identifier
239861
ISSN
1550-9087
Source
Industrial Biotechnology, 3 (1), 48-57
Document type
article