Disruption of the Gene Which Encodes a Serodiagnostic Antigen and Chitinase of the Human Fungal Pathogen Coccidioides immitis

Simple item page
dc.contributor.authorReichard, Utz
dc.contributor.authorHung, Chiung-Yu
dc.contributor.authorThomas, Pei W.
dc.contributor.authorCole, Garry T.
dc.creator.orcidhttps://orcid.org/0000-0003-1091-3420en_US
dc.date.accessioned2023-06-28T17:12:16Z
dc.date.available2023-06-28T17:12:16Z
dc.date.issued2000-10-01
dc.description.abstractDisruption of genes in medically important fungi has proved to be a powerful tool for evaluation of putative virulence factors and identification of potential protein targets for novel antifungal drugs. Chitinase has been suggested to play a pivotal role in autolysis of the parasitic cell wall of Coccidioides immitis during the asexual reproductive cycle (endosporulation) of this systemic pathogen. Two chitinase genes (CTS1 and CTS2) of C. immitis have been cloned. Preliminary evidence has suggested that expression of CTS1 is markedly increased during endospore formation. The secreted CTS1 chitinase has also been shown to react with patient anti-Coccidioides complement-fixing (CF) antibody and is a valuable aid in the serodiagnosis of coccidioidomycosis. To examine the role of CTS1 in the morphogenesis of parasitic cells, the CTS1 gene was disrupted by a single, locus-specific crossover event. This resulted in homologous integration of a pAN7.1 plasmid construct that contained a 1.1-kb fragment of the chitinase gene into the chromosomal DNA of C. immitis. Results of Southern hybridizations, immunoblot analyses of culture filtrates using both CTS1-specific murine antiserum and serum from a patient with confirmed coccidioidal infection, an immunodiffusion test for CF antigenicity, and substrate gel electrophoresis assays of chitinase activity confirmed that the CTS1 gene was disrupted and nonfunctional. This is the first report of a successful targeted gene disruption in C. immitis. However, loss ofCTS1 function had no effect on virulence or endosporulation. Comparative assays of chitinase activity in the parental and Δcts1 strains suggested that the absence of a functional CTS1 gene can be compensated for by elevated expression of the CTS2 gene. Current investigations are focused on disruption of CTS2 in the Δcts1host to further evaluate the significance of chitinase activity in the parasitic cycle of C. immitis.en_US
dc.description.departmentMolecular Microbiology and Immunologyen_US
dc.description.sponsorshipNational Institute of Allergy and Infectious Diseases, National Institutes of Healthen_US
dc.identifier.citationReichard, U., Hung, C.-Y., Thomas, P. W., & Cole, G. T. (2000). Disruption of the Gene Which Encodes a Serodiagnostic Antigen and Chitinase of the Human Fungal Pathogen Coccidioides immitis. Infection and Immunity, 68(10), 5830-5838. doi:10.1128/iai.68.10.5830-5838.2000en_US
dc.identifier.issn1098-5522
dc.identifier.otherhttps://doi.org/10.1128/iai.68.10.5830-5838.2000
dc.identifier.urihttps://hdl.handle.net/20.500.12588/1959
dc.language.isoen_USen_US
dc.publisherAmerican Society for Microbiologyen_US
dc.relation.ispartofseriesInfection and Immunity;Vol. 68, No. 10
dc.subjectCoccidioides immitisen_US
dc.subjectCoccidioidesen_US
dc.subjectcoccidioidomycosisen_US
dc.titleDisruption of the Gene Which Encodes a Serodiagnostic Antigen and Chitinase of the Human Fungal Pathogen Coccidioides immitisen_US
dc.typeArticleen_US

Files

Original bundle

Now showing 1 - 1 of 1
Thumbnail Image
Name:
Reichard 2000 - Disruption of the Gene Which Encodes a Serodiagnostic Antigen and Chitinase of the Human Fungal Pathogen Coccidioides immitis.pdf
Size:
1.66 MB
Format:
Adobe Portable Document Format
Description:
Download

License bundle

Now showing 1 - 1 of 1
No Thumbnail Available
Name:
license.txt
Size:
1.86 KB
Format:
Item-specific license agreed upon to submission
Description:
Download

Collections

College of Sciences Faculty Research