利用SSH技术分离结球甘蓝耐热性相关基因

Isolation of Resistance Related-genes to High Temperatures with Suppression Subtractive Hybridization in Cabbage

目的:找出与甘蓝耐热性相关的基因,应用抑制性差减杂交(SSH)。方法:构建了耐热结球甘蓝自交系"夏皇6-105"幼苗经高温诱导0.25h、1h、3h、6h、12h、24h后的混合SSH文库。结果:共获得300个独立克隆,经反向Northern杂交验证,对98个差异表达克隆进行了测序并利用Blastn和Blastx对测序结果进行相似性分析。结论:用此方法分离出了热激蛋白、细胞防御、信号转导、蛋白质修饰加工等与植物耐热性相关的基因片段。

Objective：To search for related-genes to high temperatures in cabbage,cDNA library of the cabbage that resistant high temperature cultivar.Method：Xiahuang 6-105,was constructed using method of suppression subtractive hybridization（SSH） under the stress of high temperature induction 0.25h,1h,3h,6h,12h and 24h.Result：The library containing 300 independent clones.The 98 differential expression clones were sequenced and sequence similarity searches were performed with the BLASTn and BLASTx.Conclusion：The fragments of genes related with the heat resistance of plant such as HSP,cell defense,signal transduction protein modification and elaboration can be separated by SSH.