优化TAIL-PCR方法克隆棉花抗逆相关转录因子编码基因  

Cloning of Stress-related Transcription Factors Gene from Cotton by Optimized TAIL-PCR

作　　者：梁成真[1] 张锐[1] 孙国清[1] 孟志刚[1] 周焘[1] 丁忠涛[1] 郭三堆[1] 

机构地区：[1]中国农业科学院生物技术研究所/国家农作物基因资源与基因改良重大科学工程,北京100081

出　　处：《棉花学报》2010年第3期195-201,共7页Cotton Science

基　　金：农业部国家转基因重大专项:"转基因抗旱耐盐碱棉花新品种培育";2008ZX005-004

摘　　要：ABF/AREB/ABI5/DPBF类转录因子属于碱性亮氨酸类蛋白,在依赖ABA的逆境信号传导途径中起重要的作用。本研究结合该类转录因子的结构特点,利用3′RACE技术和优化的TAIL-PCR技术成功获得棉花中三个该家族成员编码基因。进一步验证结果表明两步PCR成功获得基因完整的ORF、3′UTR以及部分5′UTR序列。本研究所获基因为棉花抗逆基因工程提供了候选基因源,优化后的TAIL-PCR技术为克隆棉花中目的基因提供了一种简便高效的方法。ABF/AREB/ABI5/DPBF transcription factors belong to basic leucine zipper protein,this kind of transcription factors plays an important role in the signal transduction pathways of ABA-dependent abiotic stress.In this study,taking structure char-acteristics of ABF/AREB/ABI5/DPBF as the research object,we cloned three bZIP encoding genes by 3′ RACE successfully from cotton and also optimized the TAIL-PCR conditions.The further experiment results showed that we had got the integrated sequences of the ORF,3′ UTR,as well as part of 5′ UTR sequence.The genes we cloned will provide candidate genes for resis-tance genetic engineering of cotton.Furthermore,the optimized methodology of TAIL-PCR will provide an easy and efficient technology for cloning of target genes.

关 键 词：棉花 ABF/AREB/ABI5/DPBF 转录因子 TAIL-PCR 抗逆相关基因 

分 类 号：S562[农业科学—作物学]