Fardis Teifoori1, Masoomeh Shams-Ghahfarokhi2, Mehdi Razzaghi-Abyaneh3, Jorge Martinez4. 1. Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran. 2. Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran. Electronic address: shamsm@modares.ac.ir. 3. Department of Mycology, Pasteur Institute of Iran, Tehran 13164, Iran. 4. Department of Immunology, Microbiology and Parasitology, Faculty of Pharmacy, University of the Basque Country, Vitoria, Spain.
Abstract
BACKGROUND: Members of the Pleosporaceae family are known as important sources of airborne allergens which are responsible for asthma and allergic diseases. AIMS: The purpose of this study was to investigate the gene profiling and expression pattern of Alt a 1 in Alternaria alternata and other members of the Pleosporaceae family including Stemphylium botryosum, Ulocladium chartarum, Curvularia lunata, Cladosporium cladosporioides, and Epicoccum nigrum. METHODS: Alternaria alternata and related genera were cultured on Czapek-Dox broth medium at 25°C for 21 days. The presence of Alt a 1 was assessed in fungal culture filtrates by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and then confirmed by immunoblot analysis. Real-time PCR was carried out for quantitation of the Alt a 1 gene encoding corresponding protein at the transcriptional level using cDNA prepared from fungal RNA. RESULTS: SDS-PAGE showed protein bands ranging from 14 to 100kDa. A 14kDa band corresponding to Alt a 1 was present in A. alternata, S. botryosum and U. chartarum. The gene expression of Alt a 1 was reported in A. alternata and some other related genera. The Ct mean value recorded for A. alternata strains ranged from 24.70 to 27.84 while it was in the range 23.62-32.09 for other related taxa. No apparent transcription or expression was revealed in C. cladosporioides. CONCLUSIONS: The presence and efficient expression of Alt a 1 gene in A. alternata and other related taxa indicate that Alt a 1 protein is a major component of the secretory machinery of Pleosporaceae family members, and it may play a crucial role in its allergenicity.
BACKGROUND: Members of the Pleosporaceae family are known as important sources of airborne allergens which are responsible for asthma and allergic diseases. AIMS: The purpose of this study was to investigate the gene profiling and expression pattern of Alt a 1 in Alternaria alternata and other members of the Pleosporaceae family including Stemphylium botryosum, Ulocladium chartarum, Curvularia lunata, Cladosporium cladosporioides, and Epicoccum nigrum. METHODS:Alternaria alternata and related genera were cultured on Czapek-Dox broth medium at 25°C for 21 days. The presence of Alt a 1 was assessed in fungal culture filtrates by sodium dodecyl sulfatepolyacrylamide gel electrophoresis (SDS-PAGE) and then confirmed by immunoblot analysis. Real-time PCR was carried out for quantitation of the Alt a 1 gene encoding corresponding protein at the transcriptional level using cDNA prepared from fungal RNA. RESULTS:SDS-PAGE showed protein bands ranging from 14 to 100kDa. A 14kDa band corresponding to Alt a 1 was present in A. alternata, S. botryosum and U. chartarum. The gene expression of Alt a 1 was reported in A. alternata and some other related genera. The Ct mean value recorded for A. alternata strains ranged from 24.70 to 27.84 while it was in the range 23.62-32.09 for other related taxa. No apparent transcription or expression was revealed in C. cladosporioides. CONCLUSIONS: The presence and efficient expression of Alt a 1 gene in A. alternata and other related taxa indicate that Alt a 1 protein is a major component of the secretory machinery of Pleosporaceae family members, and it may play a crucial role in its allergenicity.