摘要
采用抑制性消减杂交技术,以甘蓝型油菜矮化突变体"NDF-1"及野生型"3529"为材料构建与植株高矮性状关系最为密切的茎尖差异表达抑制消减cDNA文库。所建cDNA文库插入片段大小主要集中在100～500 bp之间。斑点杂交筛选得到30个阳性克隆,序列测定和同源性对比分析表明,所得克隆功能涉及到第二信使、转录因子、激素代谢、蛋白质降解及转运等多个方面。并对其中几个差异表达基因的功能进行了初步分析,为进一步认识油菜植株矮化机理奠定了基础。
cDNA library of differentially expressed genes in dwarf mutant＂NDF-1＂ and its wild type ＂3529＂ was constructed by suppression subtractive hybridization （SSH）. These clones were checked by colony PCR. The recombined efficiencies of the subtractive library were 58%. The size of inserted fragments was ranged from 100 to 500 bp. Eighty eight clones in the subtractive library were screened by dot blot. Thirty positive clones were sequenced and their homology was analyzed by BLAST search in GenBank. Some of these clones were put into several functional cat- egories, including signaling, secondary messenger, hormone metabolism, transcription factor, protein degradation and transfer. The successful construction of subtractive cDNA library has provided a foundation for further studying the mechanism of plant dwarfism.
出处
《河南农业科学》
CSCD
北大核心
2007年第9期33-36,共4页
Journal of Henan Agricultural Sciences
基金
国家863课题(2001AA241104)
四川省"十五"油菜育种攻关项目(200107001)
关键词
甘蓝型油菜
抑制消减杂交
CDNA文库
矮化
差异表达
Brassica napus
Suppression subtractive hybridization
cDNA library
Dwarf Mutant
Differential expression
