茶树bZIP转录因子基因CsbZIP1的克隆与表达定位.doc

茶树bZIP转录因子基因 CsbZIP1 的克隆与表达定位 曹红利 岳 川 周艳华 王 璐 郝心愿 杨亚军* 王新超 中国农业科学院茶叶研究所 / 国家茶树改良中心 / 农业部茶树生物学与资源利用重点实验室, 浙江杭州310008 * 摘 要：碱性亮氨酸拉链蛋白(bZIP)作为真核生物中分布最广、最保守的一类转录因子，参与多种生物学过程，尤其在植物 抵御各种逆境胁迫中有重要作用。采用RACE和RT-PCR技术克隆到茶树bZIP转录因子基因全长cDNA序列，命名为 CsbZIP1 (GenBank登录号为JX050148.1)。该基因cDNA全长1515 bp，包含813 bp的完整开放阅读框(ORF)，编码270个氨基 酸，预测分子量29.484 kD；含有bZIP家族典型的BRLZ结构域碱性结构域和亮氨酸拉链，属于B- 关键词：茶树；bZIP转录因子； CsbZIP1 ；亚细胞定位；克隆表达 zip1家族；系统发育树分析 显示 CsbZIP1 属于bZIP转录因子F亚家族；亚细胞定位结果表明CsbZIP1主要定位于细胞核；qRT-PCR分析表明，4低温和 NaCl盐胁迫处理均能诱导 CsbZIP1 的表达，表达量变化趋势都是随着胁迫时间先逐渐升高，到24 h时降低，ABA胁迫处理 24 h抑制 CsbZIP1 的表达。推测 CsbZIP1 与茶树低温、盐等逆境胁迫密切相关。 Molecular Cloning and Expression of a bZIP Transcription Factor Gene CsbZIP1 in Tea Plant ( Camellia sinensis ) CAO Hong-Li, YUE Chuan, ZHOU Yan-Hua, WANG Lu, HAO Xin-Yuan, YANG Ya-Jun*, and WANG Xin-Chao Tea Research Institute, Chinese Academy of Agricultural Sciences / National Center for Tea Improvement / Key Laboratory of Tea Biology and Resources Utilization, Ministry of Agriculture, Hangzhou 310008, China * Abstract: The basic leucine zipper proteins (bZIP) are one of the most extensive and conserved transcriptional factors families in eukaryotes, and plant bZIPs play important roles in many biological processes, especialy for resisting abiotic stresses. In this study, a bZIP full-length cDNA sequence was cloned using RACE and reverse transcription-PCR(RT-PCR) techniques from tea plant ( Camellia sinensis ). The obtained full-length cDNA was named CsbZIP1 with GenBank accession number JX050148.1. It is 1515 bp in length, containing a 813 bp open reading frame (ORF), encoding 270 amino acid residues with 29.484 kD molecular weight, and containing a typical BRLZ motif (basic region domain and leucine zipper domain) of B-zip1 family. The phylogenetic tree analysis revealed that CsbZIP1 belongs to F subfamily of bZIP. The subcellular location showed that CsbZIP1 protein is located in nucleus. The qRT-PCR analysis indicated that the express