Titlebook: Host-Fungus Interactions; Methods and Protocol Alexandra C. Brand,Donna M. MacCallum Book 2012 Springer Science+Business Media, LLC 2012 Fu

CANT

Mini-blaster-Mediated Targeted Gene Disruption and Marker Complementation in ,ng . gene and 200-bp flanking repeats that is useful for disruption of . genes. The cassette can be used to create unmarked homozygous mutants which can be complemented at the . gene locus. This strategy of creating gene disruptions and subsequent complementation can be used to study gene function.

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FUME

Targeted Gene Disruption in , Using Double-Joint PCR with Split Dominant Selectable Markersction of gene disruption cassettes carrying dominant selectable markers, followed by biolistic transformation. However, the conventional overlap PCR method between two flanking regions of the target gene and selectable marker is often inefficient due to the long length of the PCR product and the pre

lethal

Multiple Gene Deletion in , Using the Cre–, Systemelectable markers available for use in making gene deletions or other genetic manipulations in .. Here, we describe the adaptation of the Bacteriophage P1 Cre–. system for use in ., and its application in the excision and reuse of the geneticin drug marker. This tool will allow investigators to make

CLAP

esoteric

Targeted Gene Deletion in , Using the Hygromycin-Resistance Split-Marker Approach using site-specific recombination. Transformation of ., like many related fungal species, must overcome two major obstacles. First, the cell wall limits the entry of exogenous DNA, and second, a high rate of nonhomologous recombination leads to random ectopic integration of the marker. Here, we des

艺术

Gene Disruption in , Using Hygromycin or Phleomycin Resistance Markersarget gene of .. The DNA constructs employed contain either the gene that encodes for hygromycin B or phleomycin resistance, which are present in the pAN7.1 or pAN8.1 plasmid vectors, respectively. Hygromycin B or phleomycin are used to select for transformants at concentrations that inhibit growth

seruting

Alcove

RNA Interference in ,he process relies on double-stranded RNA (dsRNA) to target complementary messenger RNA for degradation. Here, we describe two plasmid-based strategies we have developed for RNAi in .. The pFrame vector utilizes the . promoter to enable the constitutive synthesis of hairpin RNA (hpRNA), the stem of w