优化TAIL-PCR方法克隆棉花抗逆相关转录因子编码基因.pdfVIP

棉花 学 报 Cotton Science 2010 22 渊3冤院195~201 优化TAIL-PCR 方法克隆棉花抗逆相关 转录因子编码基因 成真 张 锐 孙国清 孟志刚 周 焘 丁忠涛 郭三堆* ( 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) 摘要院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 起重 要的作用遥 本研究结合该类转录因子的结构特点 利用3忆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遥 进一步验证结果表明两步PCR 成功获得基因完整的ORF尧3忆 UTR 以及部分5忆 UTR 序 遥 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 目的基因提供了一种简便高效的方法遥 关键词院棉花曰ABF/AREB/ABI5/DPBF 曰转录因子曰TAIL-PCR 曰抗逆相关基因 中图分类号院S562.035.3 文献标识码院A 文章编号院1002-7807(2010)03-0195-07 LIANG Cheng-zhen, ZHANG Rui, SUN Guo-qing, MENG Zhi-gang, ZHOU Tao, DING Zhong-tao, GUO San-dui* ( 100081 ) 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 obj ect, we cloned three bZIP encoding genes by 3忆 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忆 UTR, as well as part of 5忆 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. cotton; ABF/AREB/ABI5/DPBF; transcription factor; TAIL-PCR; stress-related genes ABF/AREB/ABI5/DPBF 类转录因子属于 结合咱7暂 从而调节下游大量抗旱尧耐盐碱等相关功 bZIP 类蛋白 该类转录因子C 端包含一个高度 咱8暂 能基因的表达 提高植物抵御非生物逆境的能 保守的碱性亮氨酸拉链结构 N 端含有三个保 咱9-13暂