条斑紫菜cDNA微阵列制备及其在世代差异基因表达检测中的应用

为了高通量研究不同世代条斑紫菜的基因表达情况,选择了467个包含诸多功能基因的条斑紫菜基因克隆,经PCR扩增纯化后,点样于多聚赖氨酸包被的基片上制备了条斑紫菜功能基因cDNA微阵列.采用Cy3-dCTP和Cy5-dCTP 荧光分别标记条斑紫菜配子体和孢子体cRNA,与阵列进行杂交后,获得了信号清晰、重复性良好的扫描图像.经对标准化处理后的467个基因的数据进行分析发现:在配子体中表达量上调的基因有55个,其中有21个基因与已知功能基因或推测功能基因相匹配;在孢子体中表达量上调的基因有86个,其中与已知功能或假定功能基因相匹配的基因有24个.实验证明,优化的cDNA微阵列制备技术用于条斑紫菜基因表达分析有较高的效率和可靠的实用性能.

Preparation of Porphyra yezoensis cDNA microarray for analyzing the expression profile differences between sporophyte and gametophyte

To study the genes expression profiles of different generations of P. yezoensis, the eDNA mieroarray was prepared. After PCR amplification and purification, 467 P. yezoens/s functional gene clones were obtained and dotted onto the slides coated by poly-lysine. The total RNA from gametophyte and sporophyte generations sample were extracted respectively, and their cRNA were labeled by fluorescent staining. The labeled probes were then hybridized with the eDNA microarray. Uniform background and clear signal of the microarray were demonstrated by the fluorescence scanner. The analysis of the normalized data showed that 55 genes were excessive expressed in gametophyte generation, 21 of which were similar to the genes of known or putative function; 86 genes were excessive expressed in sporophyte generation, and among them, 24 were similar to the genes of known or putative function, eDNA microarray provides a high-capacity and credibility system for the investigation of Porphyra yezoensis genes expression profile.