Efficient four fragment cloning for the construction of vectors for targeted gene replacement in filamentous fungi.

ABSTRACT:

Background

The rapid increase in whole genome fungal sequence information allows large scale functional analyses of target genes. Efficient transformation methods to obtain site-directed gene replacement, targeted over-expression by promoter replacement, in-frame epitope tagging or fusion of coding sequences with fluorescent markers such as GFP are essential for this process. Construction of vectors for these experiments depends on the directional cloning of two homologous recombination sequences on each side of a selection marker gene.

Results

Here, we present a USER Friendly cloning based technique that allows single step cloning of the two required homologous recombination sequences into different sites of a recipient vector. The advantages are: A simple experimental desi

SUBMITTER: Frandsen RJ 

PROVIDER: S-EPMC2533011 | biostudies-literature | 2008 Aug

REPOSITORIES: biostudies-literature