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目的探讨申克孢子丝菌菌丝相(Mycelium,M)和酵母相(Yeast,Y)双相转换的分子机制,筛选与其双相转换相关的差异表达基因。方法应用抑制性消减杂交技术,构建申克孢子丝菌菌丝相和酵母相的正反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条非冗余序列(Unigenes);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和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 已成功构建申克孢子丝菌双相转换相关的cDNA消减文库,并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。
Objective To explore the molecular mechanism of the biphasic transformation of Mycelium (M) and Yeast (Y), and to screen the differentially expressed genes related to their biphasic transformation. Methods Suppression subtractive hybridization (SSH) was used to construct the positive and negative cDNA subtractive libraries of mycelial and yeast phases of Spirillum hypheniformis, and the differentially expressed genes were analyzed by bioinformatics methods. Results There were 751 Expressed sequence tags (ESTs) in M + Y library. 101 non-redundant sequences were obtained after splicing. 875 ESTs were obtained from Y + M library and 249 Unigenes were spliced. The transformation of Spirulina and the mycelium phase of Schenckenbachus congeners is accompanied by the high expression of differential genes in different bacterial cells. These highly expressed differential genes can be divided into structural genes, metabolic enzymes, cell surface molecules and unknown function Cell molecule. Conclusion The subtractive cDNA library of Spirulina isolates from Spirulina conidia has been successfully constructed and some differentially expressed genes were screened out, which laid the foundation of further study on the pathogenicity genes of Spirulina.