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Cloning and disruption of the antigenic catalase gene of Aspergillus fumigatus.

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TLDR
Cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain and there was no difference in pathogenicity between the cat1 mutant and its parental cat1+ strain in a murine model of aspergillosis.
Abstract
Aspergillus fumigatus possesses two catalases (described as fast and slow on the basis of their electrophoretic mobility). The slow catalase has been recognized as a diagnostic antigen for aspergillosis in immunocompetent patients. The antigenic catalase has been purified. The enzyme is a tetrameric protein composed of 90-kDa subunits. The corresponding cat1 gene was cloned, and sequencing data show that the cat1 gene codes for a 728-amino-acid polypeptide. A recombinant protein expressed in Pichia pastoris is enzymatically active and has biochemical and antigenic properties that are similar to those of the wild-type catalase. Molecular experiments reveal that CAT1 contains a signal peptide and a propeptide of 15 and 12 amino acid residues, respectively. cat1-disrupted mutants that were unable to produce the slow catalase were as sensitive to H2O2 and polymorphonuclear cells as the wild-type strain. In addition, there was no difference in pathogenicity between the cat1 mutant and its parental cat1+ strain in a murine model of aspergillosis.

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Journal ArticleDOI

Aspergillus fumigatus and Aspergillosis

TL;DR: This review focuses on the biology of A. fumigatus, one of the most ubiquitous of the airborne saprophytic fungi, and the diseases it causes, and discusses discussions of genomic and molecular characterization of the organism.
Journal ArticleDOI

Heterologous protein expression in the methylotrophic yeast Pichia pastoris

TL;DR: This paper reviews the P. pastoris expression system: how it was developed, how it works, and what proteins have been produced and describes new promoters and auxotrophic marker/host strain combinations which extend the usefulness of the system.

Aspergillus fumigatus and aspergillosis.

TL;DR: A review of the biology of Aspergillus fumigatus and the diseases it causes can be found in this article, where the authors discuss genomic and molecular characterization of the organism, clinical and laboratory methods available for the diagnosis of aspergillosis in immunocompetent and immunOCompromised hosts, identification of host and fungal factors that play a role in the establishment of the fungus in vivo, and problems associated with antifungal therapy.
Journal ArticleDOI

Recombinant protein expression in Pichia pastoris.

TL;DR: Major advances in the development of new strains and vectors, improved techniques, and the commercial availability of these tools coupled with a better understanding of the biology of Pichia species have led to this microbe’s value and power in commercial and research labs alike.
Journal ArticleDOI

Pathogenesis of Aspergillus fumigatus in Invasive Aspergillosis

TL;DR: An overview of the significant findings regarding A. fumigatus pathogenesis as it pertains to invasive disease is provided.
References
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Journal ArticleDOI

Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4

TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products.
Book

Molecular Cloning: A Laboratory Manual

TL;DR: Molecular Cloning has served as the foundation of technical expertise in labs worldwide for 30 years as mentioned in this paper and has been so popular, or so influential, that no other manual has been more widely used and influential.
Journal Article

Cleavage of structural proteins during the assemble of the head of bacterio-phage T4

U. K. Laemmli
- 01 Jan 1970 - 
TL;DR: Using an improved method of gel electrophoresis, many hitherto unknown proteins have been found in bacteriophage T4 and some of these have been identified with specific gene products as mentioned in this paper.
Journal ArticleDOI

Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications.

TL;DR: A method has been devised for the electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets that results in quantitative transfer of ribosomal proteins from gels containing urea.
Journal ArticleDOI

A new method for predicting signal sequence cleavage sites.

TL;DR: A new method for identifying secretory signal sequences and for predicting the site of cleavage between a signal sequence and the mature exported protein is described.
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