A Rapid Method for Generating Gene Deletions in Aspergillus fumigatus Protocol The technique makes use of an Escherichia coli strain expressing the redΑßΓ operon under the control of an inducible promoter. This enables the strain to carry out homologous recombination with only 50-60 bp of homologous sequence. The procedure does not require any DNA ligation and is very rapid. It allows a single gene or region on a cosmid to be replaced by a bi-functional selectable marker (having both an E. coli and an A. fumigatus marker).
Aspergillus nidulans Mutants Compilation of A. nidulans mutants listing names, affected enzymes (or gene functions), linkage assignments, properties of the mutants, and names of corresponding loci in N. crassa.
Fundamentals of Growth, Storage, Genetics and Microscopy of Aspergillus nidulans Protocols Compendium of protocols for using Aspergillus nidulans in genetic, molecular, and cell biological investigations, originally written for members of my research group. It also summarizes our common growth media and nutritional supplements, many of which originally appeared elsewhere but now are difficult to locate. Includes: Growth and storage of Aspergillus nidulans conidia; Nutritional supplements for our common auxotrophies; Double mutants; Mitotic mapping - assigning genes to chromosomes; etc
Method for Assaying Gliotoxin Production in Aspergillus fumigatus Protocol Gliotoxin is a metabolite of Aspergillus fumigatus that exhibits immunosuppressive activity against certain cells of the immune system. Secretion of gliotoxin during infection has been suggested as being a factor in the pathogenesis of aspergillosis. Gliotoxin secretion can be assayed in a number of ways by thin layer chromatography (TLC) high performance liquid chromatography (HPLC) or bioassay using the effect of gliotoxin on human cells1.
Random Amplification of Polymorphic DNA (RAPD) Typing and Fingerprinting Protocol RAPD is a procedure for typing and fingerprinting isolates of a species. It can be used for epidemiological studies, such as investigations into hospital outbreaks and as a laboratory aid to keep track of cultures and to verify that mutants generated in the laboratory are genetically identical to the parental strain. In our hands, the use of one primer, R108, is sufficiently discriminatory to distinguish between the isolates of different strains.
Sampling of Aspergillus spp Spores in Air Protocol Protocol allows the isolation and enumerate Aspergillus spp spores in air. Includes: Sampling Procedure; Sampling Location; Selection of Sampling Time; Sampling Steps; Laboratory Procedure; Enumerating the Colony Forming Units.
Transformation of Aspergillus niger Protocol Protocol for the transformation of Aspergillus niger. This procedure is done by first digesting the outer cell wall, forming protoplasts, and then by making holes in the membrane
through which the dna can enter using calcium chloride and polyethylene glycol. Includes: Protocol for making A.niger protoplasts; Transformation; Plating.
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