番茄长花柱品系T431低温胁迫下正向差减文库的构建及ESTs分析

Construction of forward subtractive library under low temperature stress on the long-style line T431of tomato and ESTs analysis

本研究以番茄长花柱品系T431为材料利用抑制性消减杂交技术(Suppression Subtractive Hybridization,SSH)构建了低温胁迫下的番茄长花柱突变体SSH cDNA正向差减文库,获得398个阳性克隆。经菌液PCR和Reverse Northern Blot杂交方法结合剔除假阳性,利用BLAST在GenBank数据库进行序列相似性比对,获得有功能的ESTs173个。通过对所获得的ESTs序列进行功能分类发现,其中参与初级代谢、能量代谢、抗病/防御反应和信号传导等过程的基因相对较多。此外,还有一些基因参与植物体内蛋白质合成与加工、物质运输、转运调控与生殖发育等过程。

Using the long-style tomato line T431 as the experimental material,we constructed a forward subtracted library by suppressive subtractive hybridization（SSH）method.The cDNA library contained 398 positive clones,which were identified through PCR and Reverse Northern Blot.By comparing with sequences in the database of GenBank with BLASTn and BLASTx algorithm,we obtained a total of 173 functional ESTs.Functional classification of these ESTs indicates that many of them are involved in primary metabolism,energy metabolism,disease resistance/defense or signal transduction.In addition,some genes are implicated in biological processes such as protein synthesis and processing,transportation regulation,and reproductive development,etc.