首页> 美国卫生研究院文献>Journal of Zhejiang University. Science. B >Construction and analysis of the cDNA subtraction library of yeast and mycelial phases of Sporothrix globosa isolated in China: identification of differentially expressed genes
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Construction and analysis of the cDNA subtraction library of yeast and mycelial phases of Sporothrix globosa isolated in China: identification of differentially expressed genes

机译:中国分离的球孢菌酵母和菌丝体相减cDNA文库的构建与分析:差异表达基因的鉴定

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摘要

Species included in the Sporothrix schenckii complex are temperature-dependent with dimorphic growth and cause sporotrichosis that is characterized by chronic and fatal lymphocutaneous lesions. The putative species included in the Sporothrix complex are S. brasiliensis, S. globosa, S. mexicana, S. pallida, S. schenckii, and S. lurei. S. globosa is the causal agent of sporotrichosis in China, and its pathogenicity appears to be closely related to the dimorphic transition, i.e. from the mycelial to the yeast phase, it adapts to changing environmental conditions. To determine the molecular mechanisms of the switching process that mediates the dimorphic transition of S. globosa, suppression subtractive hybridization (SSH) was used to prepare a complementary DNA (cDNA) subtraction library from the yeast and mycelial phases. Bioinformatics analysis was performed to profile the relationship between differently expressed genes and the dimorphic transition. Two genes that were expressed at higher levels by the yeast form were selected, and their differential expression levels were verified using a quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR). It is believed that these differently expressed genes are involved in the pathogenesis of S. globosa infection in China.
机译:包括在Sporothrix schenckii复合物中的物种是温度依赖性的,具有双态生长,并会导致孢子体增生,其特征是慢性和致命性淋巴皮肤病变。 Sporothrix复合体中包括的推定物种为巴西利亚链霉菌,球藻链球菌,墨西哥链球菌,帕氏链球菌,申氏链球菌和卢氏链球菌。球形链球菌是中国孢子菌病的病原体,其致病性似乎与双态转化密切相关,即从菌丝体到酵母相,它适应不断变化的环境条件。为了确定介导球形球菌双态转化的转换过程的分子机制,抑制消减杂交(SSH)用于制备酵母和菌丝体相的互补DNA(cDNA)扣除文库。进行了生物信息学分析,以分析不同表达的基因与双态转化之间的关系。选择通过酵母形式以较高水平表达的两个基因,并使用定量实时逆转录酶聚合酶链反应(qRT-PCR)验证其差异表达水平。相信这些不同表达的基因参与了中国的球形链球菌感染的发病机理。

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