申克孢子丝菌菌丝相和酵母相cDNA消减文库的构建及差异表达基因的筛选

目的构建申克孢子丝菌双相cDNA消减文库,筛选与双相转换相关的差异表达基因。方法运用抑制性消减杂交技术,构建申克孢子丝菌菌丝相（Mycelium,M）和酵母相（Yeast,Y）的正反cDNA消减文库,并对其差异表达的基因进行生物信息学分析。结果 M＋Y文库获得751条表达序列标签（Expressed Sequence Tags,ESTs）,经拼接后获得101条uni-genes;Y＋M文库获得875条ESTs,拼接获得249条unigenes。申克孢子丝菌酵母相菌丝相的转换伴随着不同菌相细胞差异基因的高表达,这些高表达的差异基因可分为结构基因类、代谢酶类、细胞表面分子类及功能不明的细胞分子。结论成功构建了申克孢子丝菌双相转换相关的cDNA消减文库基础上,筛选出部分差异表达基因,为进一步研究申克孢子丝菌致病相关基因奠定了基础。

Construction of cDNA subtractive library between yeast and mycelium phase of Sporothrix schenckii and screening of differently expressed genes about dimorphic transition

The purpose of this study was to screen the differentially expressed genes about dimorphic transition of Sporothrix schenckii by construction of cDNA subtractive library between yeast and mycelium phase.The cDNA subtractive library between yeast and mycelium phase was constructed by suppression subtractive hybridization（SSH）and bioinformatics analysis was performed to profile the relationship between those differently expressed genes and dimorphic transition.Of 751 and 875 ESTs were obtained in M＋Y and Y＋M library,separately.After splicing of ESTs,101 and 249 unigenes were obtained in M＋Y and Y＋M libraries.During the construction of cDNA subtractive libraries,the distribution of differently expressed genes varied with dimorphic transition.The over expressed genes with diversity and complexity were divided into structural genes,metabolic enzymes,molecule on cell surface and molecule with indistinct function.Results disclose that construction of cDNA subtractive libraries for the dimorphic transition and bioinformatics analysis for those differently expressed genes lay the foundation for screening the related genes involved in the pathogenesis of Sporothrix schenckii infection.