Laboratory of Cell Biology, Research Institute of Forestry, Chinese Academy of Forestry, Beijing 100091, PR China.
Gene. 2013 Oct 15;529(1):150-8. doi: 10.1016/j.gene.2013.07.076. Epub 2013 Aug 9.
A full-length cDNA and genomic sequences of a translationally controlled tumor protein (TCTP) gene were isolated from Japanese larch (Larix leptolepis) and designated LaTCTP. The length of the cDNA was 1, 043 bp and contained a 504 bp open reading frame that encodes a predicted protein of 167 amino acids, characterized by two signature sequences of the TCTP protein family. Analysis of the LaTCTP gene structure indicated four introns and five exons, and it is the largest of all currently known TCTP genes in plants. The 5'-flanking promoter region of LaTCTP was cloned using an improved TAIL-PCR technique. In this region we identified many important potential cis-acting elements, such as a Box-W1 (fungal elicitor responsive element), a CAT-box (cis-acting regulatory element related to meristem expression), a CGTCA-motif (cis-acting regulatory element involved in MeJA-responsiveness), a GT1-motif (light responsive element), a Skn-1-motif (cis-acting regulatory element required for endosperm expression) and a TGA-element (auxin-responsive element), suggesting that expression of LaTCTP is highly regulated. Expression analysis demonstrated ubiquitous localization of LaTCTP mRNA in the roots, stems and needles, high mRNA levels in the embryonal-suspensor mass (ESM), browning embryogenic cultures and mature somatic embryos, and low levels of mRNA at day five during somatic embryogenesis. We suggest that LaTCTP might participate in the regulation of somatic embryo development. These results provide a theoretical basis for understanding the molecular regulatory mechanism of LaTCTP and lay the foundation for artificial regulation of somatic embryogenesis.
从日本落叶松(Larix leptolepis)中分离出一个翻译控制肿瘤蛋白(TCTP)基因的全长 cDNA 和基因组序列,并将其命名为 LaTCTP。cDNA 长度为 1043bp,包含一个 504bp 的开放阅读框,编码一个由 167 个氨基酸组成的预测蛋白,其特征是具有 TCTP 蛋白家族的两个特征序列。LaTCTP 基因结构分析表明,它含有四个内含子和五个外显子,是目前所有已知植物 TCTP 基因中最大的一个。使用改良的 TAIL-PCR 技术克隆了 LaTCTP 的 5'-侧翼启动子区。在这个区域,我们鉴定了许多重要的潜在顺式作用元件,如 Box-W1(真菌诱导应答元件)、CAT-box(与分生组织表达相关的顺式作用调节元件)、CGTCA-motif(与 MeJA 反应性相关的顺式作用调节元件)、GT1-motif(光响应元件)、Skn-1-motif(胚乳表达所需的顺式作用调节元件)和 TGA-element(生长素响应元件),这表明 LaTCTP 的表达受到高度调控。表达分析表明,LaTCTP mRNA 在根、茎和针叶中广泛存在,在胚胎悬浮体(ESM)、褐化胚性培养物和成熟体细胞胚中 mRNA 水平较高,而在体细胞胚胎发生的第五天 mRNA 水平较低。我们推测 LaTCTP 可能参与调节体细胞胚胎的发育。这些结果为理解 LaTCTP 的分子调控机制提供了理论依据,并为人工调控体细胞胚胎发生奠定了基础。