一种全长cDNA抑制缩减杂交方法及其水稻诱导表达基因分离的应用  

A Full-Length cDNA Subtractive Hybridization (FLSSH) Method and Its Application in the Isolation of Inducible Expression Genes in Rice

作　　者：张群宇[1] 刘耀光[1] 

机构地区：[1]华南农业大学生命科学院,广东广州510642

出　　处：《华南农业大学学报》2010年第1期34-37,共4页Journal of South China Agricultural University

基　　金：国家自然科学基金(30470995;30740072)

摘　　要：以缩减杂交的原理和SMART cDNA合成方法为基础,开发了一种全长cDNA缩减杂交方法.该方法从微量的2个RNA样品合成全长cDNA,经PCR扩增和在两端连接不同的接头,通过2次缩减杂交获得含有高度浓缩的差异表达的全长或较大的基因片段,克隆后筛选出差异表达的基因.以噻菌灵(Probenazole,PBZ)为诱导剂处理水稻,获得了2个上调表达和1个下调表达基因的较长的cDNA片段.cDNA suppression subtractive hybridization （SSH） has been used to analyze differential gene expression widely, but there are some shortcomings in it such as difficult mRNA preparation, difficult mRNA handling and short eDNA fragments to be obtained. A Full-length eDNA SSH （FLSSH） method based on SSH and SMART eDNA synthesis method was used to obtain differentially-expressed genes of rice treated by probenazole （PBZ）. By northern blot analysis, two up-regulated and one down-regulated candidate cDNA fragments by PBZ were identified.

关 键 词：抑制缩减杂交 基因表达 噻菌灵 

分 类 号：Q754[生物学—分子生物学]