Transient marker system for iterative gene targeting of a prototrophic fungus.

ABSTRACT: Auxotrophic microorganisms are often used for genetic engineering, because their biosynthetic deficiency can be complemented by the transforming DNA and allows selection for transformants that have become prototrophic. However, when complementation is obtained by ectopic expression this may lead to unpredictable side effects on the phenotype and, consequently, misinterpretation of experimental data. There are various ways to overcome the problem of auxotrophy, but the most reliable is to restore the function of the defective biosynthetic gene at the native genomic locus. This can be done by either sexual crossing or further genetic engineering. For fungal species lacking a perfect state or situations in which gene targeting is generally cumbersome we have developed a concept that allows tr

SUBMITTER: Nielsen ML 

PROVIDER: S-EPMC2168213 | biostudies-literature | 2007 Nov

REPOSITORIES: biostudies-literature