| Autor: |
Yu, J.-J., Kirkland, T. N., Hall, L. K., Wopschall, J., Smith, R. C., Hung, C.-Y., Chen, X., Tarcha, E., Thomas, P. W., Cole, G. T. |
| Zdroj: |
Journal of Clinical Microbiology; November 2005, Vol. 43 Issue: 11 p5462-5469, 8p |
| Abstrakt: |
ABSTRACTCoccidioidesspp. (immitisand posadasii) are the causative agents of human coccidioidomycosis. In this study, we developed a novel system to overexpress coccidioidal proteins in a nonpathogenic fungus, Uncinocarpus reesii, which is closely related to Coccidioides. A promoter derived from the heat shock protein gene (HSP60) of Coccidioides posadasiiwas used to control the transcription of the inserted gene in the constructed coccidioidal protein expression vector (pCE). The chitinase gene (CTS1) of C. posadasii, which encodes the complement fixation antigen, was expressed using this system. The recombinant Cts1 protein (rCts1Ur) was induced in pCE-CTS1-transformed U. reesiiby elevating the cultivation temperature. The isolated rCts1Urshowed chitinolytic activity that was identical to that of the native protein and had serodiagnostic efficacy comparable to those of the commercially available antigens in immunodiffusion-complement fixation tests. Using the purified rCts1Ur, 74 out of the 77 coccidioidomycosis patients examined (96.1%) were positively identified by enzyme-linked immunosorbent assay. The rCts1Urprotein showed higher chitinolytic activity and slightly greater seroreactivity than the bacterially expressed recombinant Cts1. These data suggest that this novel expression system is a useful tool to produce coccidioidal antigens for use as diagnostic antigens. |
| Databáze: |
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