Unknown

Dataset Information

0

Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen.


ABSTRACT: Detection of anti-Coccidioides complement-fixing (CF) antibody is a valuable diagnostic and prognostic aid in coccidioidomycosis. The CF antibody response is directed against a heat-labile antigen that has chitinase activity, hereafter referred to as the CF/chitinase protein. To identify and clone this immunoreactive enzyme, we constructed a Coccidioides immitis cDNA lambda ZAP expression library from spherule RNA and detected fusion peptides expressing CF epitopes by immunoscreening. A cDNA clone consisting of 1,623 bp was identified, sequenced, and found to contain a single open reading frame that encodes a protein of 47 kDa with 427 amino acids. Deduced amino acid sequence analyses showed that the cloned CF/chitinase cDNA contains a 35-amino-acid region, beginning at Ser-18 and ending at and ending at Arg-52 which has 92% homology with the reported N-terminal amino acid sequence of authentic CF/chitinase protein. The first 17 amino acids in the deduced sequence of the cloned cDNA are not present on the mature CF/chitinase protein, suggesting that it may be a signal peptide. Expression of the CF/chitinase cDNA insert by using the pGEX-4T-3 vector yields a fusion peptide that bears CF-specific epitopes and shows chitinase activity. The CF/chitinase clone will enable large-scale production of the recombinant CF antigen for use in immunoassays and facilitate studies on the role of chitinase in the morphogenesis of C. immitis.

SUBMITTER: Yang C 

PROVIDER: S-EPMC174027 | biostudies-other | 1996 Jun

REPOSITORIES: biostudies-other

altmetric image

Publications

Molecular cloning and characterization of the Coccidioides immitis complement fixation/chitinase antigen.

Yang C C   Zhu Y Y   Magee D M DM   Cox R A RA  

Infection and immunity 19960601 6


Detection of anti-Coccidioides complement-fixing (CF) antibody is a valuable diagnostic and prognostic aid in coccidioidomycosis. The CF antibody response is directed against a heat-labile antigen that has chitinase activity, hereafter referred to as the CF/chitinase protein. To identify and clone this immunoreactive enzyme, we constructed a Coccidioides immitis cDNA lambda ZAP expression library from spherule RNA and detected fusion peptides expressing CF epitopes by immunoscreening. A cDNA clo  ...[more]

Similar Datasets

| S-EPMC174476 | biostudies-other
| S-EPMC229475 | biostudies-other
| S-EPMC174128 | biostudies-other
| S-EPMC173561 | biostudies-other
| S-EPMC1287831 | biostudies-literature
| S-EPMC101544 | biostudies-literature
| S-EPMC98148 | biostudies-literature
| S-EPMC2144563 | biostudies-other
| PRJNA820037 | ENA
| PRJNA12821 | ENA