大蕉冷诱导差减文库的构建与分析

以低温处理的大蕉幼苗cDNA为检测子,未处理的大蕉幼苗cDNA为驱赶子,利用抑制性差减杂交技术(suppression subtractive hybridization,SSH)构建了大蕉幼苗冷诱导差减cDNA文库,通过点杂交差异筛选cD-NA文库,得到约50个低温下表达增强的候选克隆,对其进行DNA测序和同源性比较,发现获得的ESTs在功能上主要涉及信号传导、表达调控、非生物胁迫防御、蛋白质加工和能量代谢等方面。该SSH文库的构建为克隆大蕉抗寒相关基因奠定了基础。

Construction and Analysis of Plantain (Musa paradisiaca L.,ABB group)Seedlings Suppression Subtractive Hybridization Library During Cold Stress

To investigate cold induced genes of plantain (Musa paradisiaca L.,ABB group） seedlings, a forward subtractive cDNA library is constructed by suppression subtractive hybridization (SSH) method, which is performed using the cDNA from plantain seedlings treated with low temperature as tester and those from untreated seedlings as driver.The library is differently screened through dot hybridization, about fifty clones are identified as cold specifically induced or highly expressed. After sequencing and bioinformatics analysis, the results show that these expressed sequence tags （ESTs） are mainly related to genes involved in cell structure, protein synthesis, expression regulation, signal transduction and other uncertain functions. The research has established a basis for studying low temperature tolerance mechanism of plantain and further cloning genes related to cold resistance.