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 100 kDa. A 14 kDa 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_t 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.

中文翻译：

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交替链格孢菌和炭疽菌科相关成员中主要变应原Alt a 1的基因分析和表达。

背景

伞形科的成员被认为是导致哮喘和过敏性疾病的空气中过敏原的重要来源。

目的

本研究的目的是调查在Alt键1的基因谱分析和表达模式链格孢和Pleosporaceae家族的其他成员，包括匍柄botryosum，是Ulocladium chartarum，新月弯孢霉，芽枝状枝孢，和附球菌葵。

方法

链格孢和相关属在25℃的Czapek–Dox肉汤培养基上培养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，然后通过免疫印迹分析进行确认。使用从真菌RNA制备的cDNA，在转录水平上进行实时PCR定量编码相应蛋白质的Alt a 1基因。

结果

SDS-PAGE显示蛋白质条带范围为14至100 kDa。对应于Alt键1 A 14 kDa的带存在于链格孢，S. botryosum和U. chartarum。交替链霉菌和其他一些相关属中报道了Alt a 1的基因表达。所述Ç_吨记录平均值A.孢菌株从24.70到不等27.84，而这是在其他相关类群的范围23.62-32.09。没有明显的转录或表达被发现在C. cladosporioides。

结论

交替链霉菌和其他相关类群中Alt a 1基因的存在和有效表达表明Alt a 1蛋白是古菌科成员分泌机制的主要组成部分，并且可能在其致敏性中起关键作用。