Unknown

Dataset Information

0

Molecular cloning and characterization of Coccidioides immitis antigen 2 cDNA.


ABSTRACT: Previous experiments have provided evidence that Coccidioides immitis antigen 2 (Ag2) is a major T-cell-reactive component of mycelia and spherule cell walls. Here we report the identification and cloning of the cDNA that encodes Ag2 from a lambda ZAP cDNA expression library constructed from spherule-derived RNA. DNA sequence analysis established that the 1,255-bp clone contains a 174-bp 5' untranslated region, a 582-bp open reading frame which encodes for a protein consisting of 194 amino acids, and a 375-bp 3' untranslated region, including a poly(A) tail. The recombinant Ag2 protein has a predicted molecular mass of 19.5 kDa and contains an 18-amino-acid N terminus which has been tentatively identified as a signal peptide. The Ag2 cDNA was ligated into the pGEX-4T-3 vector and expressed in Escherichia coli TG-1 cells as a glutathione S-transferase fusion protein. The recombinant fusion protein showed reactivity with sera from patients with coccidioidomycosis and elicited delayed-type footpad hypersensitivity responses in Coccidioides-immune mice. These results suggest that the Ag2 cDNA can be used for the large-scale production of this immunologically important protein.

SUBMITTER: Zhu Y 

PROVIDER: S-EPMC174128 | biostudies-other | 1996 Jul

REPOSITORIES: biostudies-other

altmetric image

Publications

Molecular cloning and characterization of Coccidioides immitis antigen 2 cDNA.

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

Infection and immunity 19960701 7


Previous experiments have provided evidence that Coccidioides immitis antigen 2 (Ag2) is a major T-cell-reactive component of mycelia and spherule cell walls. Here we report the identification and cloning of the cDNA that encodes Ag2 from a lambda ZAP cDNA expression library constructed from spherule-derived RNA. DNA sequence analysis established that the 1,255-bp clone contains a 174-bp 5' untranslated region, a 582-bp open reading frame which encodes for a protein consisting of 194 amino acids  ...[more]

Similar Datasets

| S-EPMC174027 | biostudies-other
| S-EPMC173561 | biostudies-other
| S-EPMC174476 | biostudies-other
| S-EPMC98148 | biostudies-literature
| PRJNA12821 | ENA
| PRJNA820037 | ENA
| PRJNA245906 | ENA
| S-EPMC8496281 | biostudies-literature
| S-EPMC308958 | biostudies-literature
| S-EPMC5029449 | biostudies-literature