Advancing USER cloning into simpleUSER and nicking cloning.

The two novel methods for DNA cloning presented here have been developed for the rapid construction of vectors used for insertion of genes in filamentous fungi. The current study shows that both simpleUSER cloning and nicking cloning can substitute USER cloning for insertion of single PCR fragments into plasmids. The simpleUSER cloning method proposed in this paper varies from USER cloning by substituting the dual enzymatic plasmid preparation step with a single enzymatic step.

PHUSER (Primer Help for USER): a novel tool for USER fusion primer design.

Uracil-Specific Exision Reagent (USER) fusion is a recently developed technique that allows for assembly of multiple DNA fragments in a few simple steps. However, designing primers for USER fusion is both tedious and time consuming. Here, we present the Primer Help for USER (PHUSER) software, a novel tool for designing primers specifically for USER fusion and USER cloning applications.

Versatile enzyme expression and characterization system for Aspergillus nidulans, with the Penicillium brevicompactum polyketide synthase gene from the mycophenolic acid gene cluster as a test case.

Assigning functions to newly discovered genes constitutes one of the major challenges en route to fully exploiting the data becoming available from the genome sequencing initiatives. Heterologous expression in an appropriate host is central in functional genomics studies. In this context, filamentous fungi offer many advantages over bacterial and yeast systems.