AtDREB2B基因的克隆及植物表达载体的构建  

Gene Cloning of AtDREB2B and Constructtion of Its Plant Expression Vectors

作　　者：王天祥[1] 严晓丹[1] 贾文锁[2] 王华芳[1] 

机构地区：[1]北京林业大学生物科学与技术学院,北京100083 [2]中国农业大学农学与生物技术学院,北京100094

出　　处：《安徽农业科学》2008年第21期8943-8945,共3页Journal of Anhui Agricultural Sciences

基　　金：国家林业局948项目(2007-4-02);国家林业局公益林研究项目(200704017)

摘　　要：[目的]获得耐旱转录因子的植物表达载体,进而转化植物获得耐旱性提高的新植物株系。[方法]提取脱水处理的拟南芥总RNA,利用AtDREB2B特异引物通过RT-PCR扩增出1.2kb片段,并将其克隆至pBS-T上,测序。[结果]序列分析表明,该克隆序列与GenBank上登录AtDREB2B基因序列的一致性为100%。进一步通过片段的亚克隆,将其构建至植物表达载体pCAMBIA1301和pBin438上,分别由rd29A启动子和重复35S启动子调控基因表达。[结论]成功构建了2种AtDREB2B的植物表达载体,并通过冻融法转化根癌农杆菌GV3101,为农杆菌介导的AtDREB2B基因对植物的遗传转化奠定基础。[Objective] The research aimed to obtain the plant drought-resistant transcription factor and its expression vector,then the new plant strains of drought-resistant plants.[Method] Total RNA was obtained from dehydrated Arabidopsis.And the AtDREB2B special primers were used to amplify 1.2 kb fragment by RT-PCR.This 1.2 kb fragment was inserted into pBS-T and sequenced.[Result] Sequence analysis showed that the sequence cloned in this experiment had a similarity of 100% with AtDREB2B gene logged on GenBank.Furthermore the subcloned fragment was inserted into plant expression vectors pCAMBIA1301 and pBin438 b,which were expressed by rd29A promoter and repeated 35 S promoter regulated gene separately.[Conclusion] 2 plant expression vectors of AtDREB2B was constructed successfully.And a basic way for using Agrobacterium mediated AtDREB2B gene transformation was provided with the freezing-melting transformation of Agrobacterium GV3101.

关 键 词：DREB2B 载体构建 基因克隆 不完全酶切 

分 类 号：S188[农业科学—农业基础科学]