应用差减杂交筛选雪莲低温特异表达基因  

Isolating Cold-Regulated Genes from Sasussured involucrata Kar Seedlings through Suppression Subtractive Hybridization

作　　者：王博[1] 艾秀莲[1] 王志方[1] 李芳[2] 罗明[2] 张学军[2] 

机构地区：[1]新疆农业科学院微生物应用研究所,新疆乌鲁木齐830091 [2]新疆农业大学,新疆乌鲁木齐830091

出　　处：《生物技术》2007年第4期45-48,共4页Biotechnology

基　　金：国家自然科学基金项目资助(编号:30160039)

摘　　要：目的:获得雪莲低温特异表达基因cDNA全长片段。方法:以低温诱导雪莲为目的材料,常温诱导为对照,采用抑制差减杂交(SSH)方法,构建了雪莲的差减cDNA文库,筛选文库并得到38个cDNA片段。结果:利用GenBank数据库Blastx进行同源性搜索发现有22个非冗余克隆,其中18个已知基因4个未知基因。结论:同源分析表明冷诱导基因和磷脂酰乙醇胺结合蛋白(PEBP)也在其中。所取得的结果为日后进一步对新疆雪莲抗寒机理的研究和更好地分离低温特异表达基因奠定了基础。Objective：To acquire the full length of Cold- Regulated Genes from Sasussured involucrata Kar.Methods： Suppression subtraetive hybridization（SSH）was utilized to identify the host genes that were differentially expressed in the Cold - Regulated Genes from Sasussured involucrata Kar. In vitro cultured shoots inoculated with cold- induced Sasussured involucrata Kar were used as ＂tester＂and normal temperature- induced as ＂driver＂. Results： Subtractive cDNA library was constructed, which enriched for resistance - related genes to Sasussured. Eighty---one cDNA fragments, The Blastx homology search analysis revealed the 22 nonredundant clones were classified into the following groups： 18 known function genes,4 of unknown function.Conclusion： Homology analysis showed that cold regulated gene and phosphatidyl ethanolamine- binding family protein（PEBP）was among them. The results are good enough for further cloning of the Cold - Regulated specific expression genes and mechanism of chilling- resistance adds much genetics information of Saussurea involucrata Kar.

关 键 词：抑制差减杂交 新疆雪莲 特异表达基因 

分 类 号：Q781[生物学—分子生物学]