Gateway技术构建盐和干旱胁迫下水稻根系混合cDNA文库及其质量鉴定  

Construction and identification of a mixed cDNA library of rice roots under salt and drought stress using Gateway technology

作　　者：彭晓珏[1] 刘瑢[1] 阳菁[2,3] 丁霞[1] 王芳[1] 李阳生[2,3] 蔡耀辉 

机构地区：[1]南昌大学生命科学与食品工程学院/分子生物学与基因工程实验室,南昌330031 [2]武汉大学生命科学学院/植物遗传发育学教育部重点实验室,武汉430072 [3]杂交水稻国家重点实验,武汉430072 [4]江西省超级水稻研究发展中心,南昌330200

出　　处：《应用与环境生物学报》2014年第2期291-294,共4页Chinese Journal of Applied and Environmental Biology

基　　金：国家科学自然基金项目(31160270;31160019);教育部科学研究重大项目(313039);江西省青年科学基金项目(20114BAB-214009)资助~~

摘　　要：为研究水稻非生物胁迫响应信号转导的网络途径,揭示水稻抗逆的分子机制,以NaCl胁迫、干旱胁迫处理及正常生长的水稻根系为材料,采用Gateway技术构建了水稻根系受盐和干旱诱导的混合cDNA文库.cDNA文库质量分析表明,未经扩增的原始文库容量为1.5×106 cfu,插入片段大小主要为1 kb左右,重组率为100%.文库随机挑选克隆测序,结果显示插入序列完整性良好,文库中包含水解酶基因、逆境响应蛋白基因、氧化还原酶基因和富含甘氨酸的RNA结合蛋白质基因等.因此,本研究构建的盐和干旱诱导的水稻根系混合cDNA文库符合高质量文库的标准,可以用于进一步开展相关基因的克隆及功能研究.In order to study the pathway of rice response to abiotic stresses and discover the molecular mechanisms of stress tolerance in rice, we constructed a mixed cDNA library by using Gateway technology from rice roots subjected to salt and drought treatments. The quality assessment of this library showed that the titer of the unamplified library was 1.5 x 106 cFH, and the average insert size about 1 kb with recombination efficiency of 100%. In this study, randomly-selected clones were sequenced to find many functional genes, including glycosyl hydrolases, stress induced protein, oxidoreductase and glycine- rich RNA-binding protein, etc. The results suggested that the mixed cDNA library from rice roots was successfully generated in high quality, and it may provide an essential resource for further stress-responsive gene cloning and functional study.

关 键 词：水稻 抗逆 GATEWAY CDNA文库 盐胁迫 干旱胁迫 

分 类 号：S511[农业科学—作物学]