白念珠菌酵母相细胞长片段基因表达系列分析文库的构建和初步分析

Long serial analysis of gene expression in the yeast phase of Candida albicans

目的构建白念珠菌酵母相细胞长片段基因表达系列分析文库并探讨其基因表达特点。方法采用长片段基因表达系列分析技术(LongSAGE)构建白念珠菌酵母相细胞LongSAGE文库,并从中挑取1000个阳性克隆进行测序。结果成功构建了白念珠菌酵母相细胞LongSAGE文库。1000个测序文件中,共获得标签17773个,代表单一转录本7333种,其中单一匹配标签占16·65%,多重匹配标签占6·59%,推测蛋白标签占16·27%,基因组及粘粒等标签占1·64%,新标签为58·86%。标签拷贝数超过35的33个高表达标签中有11个可能属于新的表达序列。结论LongSAGE是一种能够快速、高通量地研究细胞基因表达谱及其丰度的方法,可发现新的表达序列。

Objective To construct a library of long serial analysis of gene expression （LongSAGE） and investigate the characteristics of gene expression in the yeast phase of C. albicans. Methods A LongSAGE library was constructed with long serial analysis of gene expression （LongSAGE）. Results A total of 17 773 tags, representing specific 7 333 transcripts, were extracted from 1 000 sequence files in the yeast phase of C. albicans. Tags matching single gene accounted for 16. 65%, tags matching multiple genes accounted for 6. 59%, tags for hypothetical protein accounted for 16. 27%, tags for genome and cosmid accounted for 1.64%, and novel tags accounted for 58. 86%. Eleven tags in 33 tags whose expression frequencies exceeded 35 may be new expression sequences. Conclusion LongSAGE is not only a powerful method in investigating the gene expression profiling, allowing the analysis of the expression of thousands of transcripts in a quantitative manner without prior sequence information, but also in finding new genes.