香蕉叶片低温胁迫下差异表达cDNA消减文库的构建  

Construction of cDNA Subtractive Library of Banana Leaf under Low Temperature Stress

作　　者：祁君凤[1] 冯仁军[2] 程萍[2] 袁克华[2] 张丽丽[2] 张银东[1,2] 

机构地区：[1]海南大学农学院,海南海口570228 [2]中国热带农业科学院热带生物技术研究所农业部热带作物生物技术重点开放实验室,海南海口571101

出　　处：《热带生物学报》2010年第2期110-113,共4页Journal of Tropical Biology

摘　　要：为了解析香蕉寒害的分子机理,应用抑制性消减杂交技术(suppression subtractivehy bridization,SSH)成功构建了香蕉叶片在低温胁迫下与未处理的差异表达的cDNA消减文库,并通过经蓝、白斑筛选,克隆测序及序列分析,以及应用半定量PCR技术对随机选取的No.28基因片段进行表达分析等方法,初步分析了实验所构建的cDNA消减文库。结果表明,实验所构建的cDNA消减文库为低温胁迫特异表达的cDNA消减文库;测序及序列分析的结果暗示可能有相当多的未知功能基因参与了香蕉抗寒的过程;随机选取的No.28的表达分析检测结果证实,通过对构建的cDNA消减文库的全面分析,有可能获得一批低温胁迫诱导特异表达的新基因,尤其对其功能的深入了解,将为全面解析香蕉寒害的分子机理、寒害过程中的信号转导打下理论基础。To elucidate the molecular mechanism of cold damage to banana, suppression subtractive hybridization （SSH） library was successfully constructed under low temperature stress and the natural growth conditions. The library was analyzed by the blue ＆ white spot screening, cloning, sequencing and semi-qPCR for No. 28, and the results indicated that there may be some unknown gene involving the anti-cold of banana except No. 28, and it is very possible to obtain more specific express gene under low temperature stress.

关 键 词：香蕉叶片 低温胁迫 SSH差减文库 

分 类 号：Q786[生物学—分子生物学]