申克孢子丝菌酵母相和菌丝相cDNA消减文库的构建  

Construction of cDNA subtractive library for the yeast and mycelium phases of Sporothrix Schenckii

作　　者：周汛[1] 杨致邦[2] 郭丽媛[2] 肖异珠[3] 

机构地区：[1]重庆医科大学附属第一医院皮肤科,重庆市400016 [2]重庆医科大学基础医学实验教学中心病原生物学与免疫学实验室,重庆市400016 [3]重庆医科大学附属儿童医院皮肤科,重庆市400014

出　　处：《医学分子生物学杂志》2011年第2期132-137,共6页Journal of Medical Molecular Biology

基　　金：重庆市卫生局科学基金（No.2008-2-73）

摘　　要：目的 筛选与申克孢子丝菌酵母相与菌丝相双相转换相关的差异表达基因,为探讨其双相转换的分子的机制奠定基础.方法 应用抑制性消减杂交技术,构建高特异性的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,平均长度为690.98 bp,经拼接后获得101条非冗余序列.Y＋M文库获得875条表达序列标签,平均长度为575.9 bp,拼接获得249条非冗余序列.经BLASTN比对,这些差异表达基因中,某些结构基因和功能不明的细胞分子类基因可能与双相转换有关.结论 成功构建了高特异性的申克孢子丝菌菌丝相和酵母相的正反cDNA 消减文库,为进一步筛选申克孢子丝菌的双相转换基因奠定了基础.Objective To explore molecular mechanism of dimorphic transition of Sporothrix schenckii, differential gene expression in dimorphic transition of Sporothrix sehenckii was screened. Methods cDNA subtractive library for the yeast （Y） and myeelium （M） phases was constructed by suppression subtractive hybridization （SSH） . Bioinformaties analysis was performed to profile the relationship between the differently expressed genes and dimorphic transition. Results 751 ESTs were obtained in M ＋ Y library, with the average length of 690. 98bp. Meanwhile, 875 ESTs were obtained in Y ＋ M library, with the average length of 575.9bp. After splicing of ESTs, 101 unigenes were obtained in M ＋ Y library and 249 unigenes in Y ＋ M library. Some structure genes and function-unknown genes in these differentially expressed genes appeared to be related to dimorphic transition as compared with BLASTN. Conclusion It is evident that the subtracted cDNA library for the dimorphic transition of Sporothrix Schenckii was successfully constructed, thereby providing solid foundation for screening the genes involved the dimorphic transition of Sporothrix schenckii.

关 键 词：申克孢子丝菌 抑制性消减杂交 cDNA 消减文库 

分 类 号：R379.2[医药卫生—病原生物学]