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Published on: February 25, 2010
Isolation and expression of heat shock protein 30 gene from Penicillium marneffei
Nongnuch Vanittanakom1, Monsicha Pongpom, Jutarat Praparattanapan
1Department of Microbiology, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand. nvanitta@mail.med.cmu.ac.th
Abstract:
Penicillium marneffei is a dimorphic fungus that can cause disseminated mycosis, especially in AIDS patients. The role of heat shock proteins and stress response-related proteins in P. marneffei remains unknown. In this study, we isolated a cDNA encoding for heat shock protein 30 (Hsp30) of P. marneffei using an antibody screening method. The DNA sequence and deduced amino acid sequence analysis showed high homology to other fungal hsp30 genes. Expression of P. marneffei hsp30 in response to temperature increase was determined by Northern blot analysis. A high level of hsp30 transcript was detected in yeast cells grown at 37 degrees C, whereas a very low or undetectable transcript level was observed in mycelial cells at 25 degrees C. A recombinant Hsp30 protein was produced and tested preliminarily for its immunoreactivity with sera from P. marneffei-infected AIDS patients using Western blot analysis. The positive immunoblot result, with some serum samples, confirmed the antigenic property of the Hsp30. Collectively, the high response of hsp30 to temperature increase could indicate it may play a role in heat stress response and cell adaptation. This is the first report showing that this small heat shock protein could elicit the human immune response.
Insights
Penicillium marneffei heat shock protein 30 (Hsp30) is upregulated in yeast cells at 37°C. This Hsp30 shows antigenic properties and may aid fungal adaptation to heat stress.
Area of Science:
- Medical Mycology
- Molecular Biology
- Immunology
Background:
- Penicillium marneffei is a dimorphic fungus causing disseminated mycosis, particularly in immunocompromised individuals.
- The function of heat shock proteins (HSPs) and stress response proteins in P. marneffei is not well understood.
Purpose of the Study:
- To isolate and characterize the heat shock protein 30 (Hsp30) gene from P. marneffei.
- To investigate the expression pattern of P. marneffei Hsp30 under thermal stress.
- To evaluate the potential immunogenicity of P. marneffei Hsp30.
Main Methods:
- Antibody screening was used to isolate the P. marneffei hsp30 cDNA.
- Northern blot analysis was performed to assess hsp30 gene expression at different temperatures (25°C and 37°C).
- Recombinant Hsp30 protein was produced and tested for immunoreactivity using Western blot analysis with patient sera.
Main Results:
- The isolated P. marneffei hsp30 cDNA showed high homology to other fungal hsp30 genes.
- hsp30 transcript levels were significantly higher in yeast cells at 37°C compared to mycelial cells at 25°C.
- Western blot analysis confirmed the antigenic properties of the recombinant Hsp30 protein, with positive reactions observed with some sera from infected patients.
Conclusions:
- P. marneffei Hsp30 expression is temperature-dependent, suggesting a role in heat stress response and cellular adaptation.
- This study is the first to demonstrate that P. marneffei Hsp30 can elicit a human immune response, highlighting its potential as an antigen.
