Cloning of a Gene Encoding an Alt a 1 Isoallergen Differentially Expressed by the Necrotrophic Fungus Alternaria brassicicola during Arabidopsis Infection
Open accessAbstract
Alternaria species are considered some of the most important fungi responsible for allergenic morbidity in humans. The Alternaria protein that elicits the most intense allergic reaction in humans is Alt a 1, yet no biological function has been identified for this protein. In this study, suppression subtractive hybridization and virtual Northern blots were used to identify and characterize an Alt a 1 homolog in the phytopathogenic fungus Alternaria brassicicola. RNA was extracted from A. brassicicola spores germinated in water and on leaf surfaces of the Arabidopsis ecotype Landsberg for 24 h and used to create cDNA by PCR. Double-stranded cDNA was then used in suppression subtractive hybridization to identify differentially expressed genes. mRNA transcript levels were assessed by virtual Northern blotting. A sequence with significant homology (90% amino acid, 92% cDNA) to the Alt a 1 subunit from Alternaria alternata was identified. Virtual Northern blots demonstrated that this homolog, designated Alt b 1 precursor, was highly up-regulated during the infection process of A. brassicicola on Arabidopsis: The full-length cDNA sequence of Alt b 1 was 815 bp, with an open reading frame of 477 bp. In this preliminary study, we identified a homolog of the major Alternaria allergen precursor, Alt a 1, in A. brassicicola, designated Alt b 1. This isoallergen is differentially expressed during fungal pathogenesis on Arabidopsis, suggesting a possible biological role in pathogenesis.
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Homologous recombination at the srbA locus in transformants was confirmed by Southern analysis.
“Homologous recombination was confirmed by Southern analysis with the digoxigenin labeling system (Roche Molecular Biochemicals, Mannheim, Germany) as previously described [71].”
From A Sterol-Regulatory Element Binding Protein Is Required for Cell Polarity, Hypoxia Adaptation, Azole Drug Resistance, and Virulence in Aspergillus fumigatus (2008) · cites this paper as [71]
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- Authors
- Cramer, R. A., & Lawrence, C. B.
- Affiliations
- Colorado State University (US)
- Corresponding author
- Lawrence, Christopher B.
- Venue
- Applied and Environmental Microbiology
- Publisher
- American Society for Microbiology
- Published
- 2003-04
- Volume
- 69
- Issue
- 4
- Pages
- 2361-2364
- ISSN
- 0099-2240 (ISSN-L)
- 1098-5336
- PMC
- PMC154767
- OpenAlex
- W2155943552
- Cited by
- 41
- References
- 14
- Citations by year
- 2026: 2 · 2024: 1 · 2022: 2 · 2021: 1 · 2020: 1 · 2019: 3 · 2018: 2 · 2017: 2 · 2016: 1 · 2015: 2 · 2014: 2 · 2012: 2
- FWCI
- 0.73
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Applied and Environmental Microbiology (publishedVersion)
- Europe PMC (PubMed Central) (publishedVersion)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Plant Pathogens and Fungal Diseases
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
- Life Sciences › Agricultural and Biological Sciences › Plant Science › Plant-Microbe Interactions and Immunity
- Keywords
- Alt a 1, Alternaria brassicicola, isoallergens, differentially expressed genes, fungal pathogenesis, Arabidopsis thaliana, suppression subtractive hybridization, cDNA cloning, necrotrophic fungi, plant-fungal interactions
- MeSH terms
- Allergens, Alternaria, Amino Acid Sequence, Antigens, Fungal, Base Sequence, Cloning, Molecular, Fungal Proteins, Molecular Sequence Data, Plant Diseases, Spores, Fungal, Gene Expression Regulation, Fungal, Arabidopsis, Sequence Analysis, DNA, DNA, Complementary, Plant Leaves, Antigens, Plant
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