cDNA cloning, genomic organization and expression analysis during somatic embryogenesis of the translationally controlled tumor protein (TCTP) gene from Japanese larch (Larix leptolepis)

• Published in: Gene 2013-10, Vol.529 (1), p.150-158
• Main Authors: Zhang, Li-feng, Li, Wan-feng, Han, Su-ying, Yang, Wen-hua, Qi, Li-wang
• Format: Article
• Language: English
• Subjects: Amino Acid Sequence; amino acids; Base Sequence; Cloning, Molecular; complementary DNA; DNA, Complementary - genetics; elicitors; endosperm; exons; fungi; gene expression; Gene Expression Profiling; Gene Expression Regulation, Plant; genes; Genomic organization; introns; Larix - genetics; Larix kaempferi; Larix leptolepis; meristems; messenger RNA; Molecular Sequence Data; Open Reading Frames; Phylogeny; Plant Proteins - genetics; Plant Proteins - metabolism; Plant Somatic Embryogenesis Techniques; Polymerase Chain Reaction; promoter regions; Promoter Regions, Genetic; Protein Conformation; Real-time quantitative PCR; RNA, Messenger - genetics; RNA, Messenger - metabolism; roots; Sequence Analysis, DNA; Somatic embryogenesis; somatic embryos; stems; Translationally controlled tumor protein
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/j.gene.2013.07.076

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 1043bp and contained a 504bp 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. •The full-length cDNA of LaTCTP was cloned and identified for the first time.•The 5′-flanking promoter of LaTCTP was cloned using an improved TAIL-PCR technique.•The 3D-structure of LaTCTP was predicted using the I-TASSER server.•The expression profiles of LaTCTP during somatic embryogenesis were presented.•LaTCTP might participate in the regulation of somatic embryo development.