Teifoori Fardis, Shams-Ghahfarokhi Masoomeh, Razzaghi-Abyaneh Mehdi, Martinez Jorge
Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran.
Department of Mycology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran 14115-331, Iran.
Rev Iberoam Micol. 2019 Apr-Jun;36(2):66-71. doi: 10.1016/j.riam.2018.01.006.
Members of the Pleosporaceae family are known as important sources of airborne allergens which are responsible for asthma and allergic diseases.
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.
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.
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 C 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.
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.
格孢腔菌科成员是空气传播变应原的重要来源，可引发哮喘和过敏性疾病。
本研究旨在调查链格孢及格孢腔菌科其他成员（包括匍柄霉、炭疽菌、新月弯孢霉、枝孢霉和黑附球菌）中Alt a 1的基因图谱和表达模式。
将链格孢及相关属种在查氏肉汤培养基上于25°C培养21天。通过十二烷基硫酸钠聚丙烯酰胺凝胶电泳（SDS-PAGE）评估真菌培养滤液中Alt a 1的存在情况，然后通过免疫印迹分析进行确认。使用从真菌RNA制备的cDNA，通过实时PCR在转录水平定量编码相应蛋白质的Alt a 1基因。
SDS-PAGE显示出分子量在14至100kDa之间的蛋白条带。链格孢、匍柄霉和炭疽菌中存在一条对应于Alt a 1的14kDa条带。在链格孢和其他一些相关属中报道了Alt a 1的基因表达。链格孢菌株的C平均值在24.70至27.84之间，而其他相关分类群的C平均值在23.62 - 32.09之间。枝孢霉中未显示出明显的转录或表达。
Alt a 1基因在链格孢和其他相关分类群中的存在及高效表达表明，Alt a 1蛋白是格孢腔菌科成员分泌机制的主要成分，并且它可能在其变应原性中起关键作用。
