差异展示法鉴定GA_3诱导的水稻差异表达的mRNA

Identification of differentially expressed mRNA species in rice(Oryza sativa) induced by GA_3 using a differential display technique

赤霉素（ｇｉｂｂｅｒｅｌｌｉｎｓ）是植物生长发育过程中一类重要的调节激素，在水稻不育系上喷施合适浓度的ＧＡ３（赤霉素的一种）可以克服其包颈现象．运用反转录和聚合酶链式反应（ＰＣＲ）建立了一套旨在分离差异表达。ＤＮＡ的差异展示方法．将赤霉素ＧＡ３处理后的苗期水稻分别制备总ｍＲＮＡ，反转录ＰＣＲ后在测序胶上比较各处理间基因表达的差异，发现其中两个基因的表达量随ＧＡ３浓度的增加而增强，另有一个基因的表达则随ＧＡ３浓度的增加而减弱．已从测序胶中回收前两个表达增强的基因的。ＤＮＡ片段并重新扩增，拟用作探针从水稻基因组文库中分离这两个受ＧＡ３调控的基因．

ibberellins(GAs) is one group of the most important plant growth regulators. A differential display scheme for the isolation of differentially expressed cDNA was established,involving reverse transcription of total mRNA and polymerase chain reaction amplification of the cDNA subpopulation. The changes of gene expression in rice seedlings affected by GA3 were analyzed through this scheme. On sequencing gels it was found that two bands, which were subsequently recovered from gels and reamplified,appeared while one band disappeared with the increase in GA3 concentration. It is suggested that these differentially expressed genes can be isolated from certain genomic library with these amplified cDNA fragments as probes.