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| Cloning and Expression Analysis of trim36 Gene in Half-smooth Tongue Sole (Cynoglossus semilaevis) |
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Abstract The tripartite motif-containing protein (TRIM) family belongs to a E3 ubiquitin protein ligase with conserved structure and rapid evolution. They are involved in a variety of biological processes, such as cell regulation, cell differentiation, metabolism and innate immunity. TRIM36 is one of members of TRIM family. In this study, trim36 gene (GenBank No. KU244470) was RACE cloned in the half-smooth tongue sole (Cynoglossus semilaevis) based on the entire genome and transcriptome sequence. trim36 gene contained 2 899 bp, and the ORF was 2 214 bp, which encoded 737 amino acids, 289 bp of 5'-untranslated regions (UTR) and 396 bp of 3'-UTR. The bioinformatics analysis showed that the amino acid sequences contained a RING finger, 2 B-Box-type zinc finger (BBOX), a coiled-coil motif, a fibronectin type Ⅲ(FN3) and spla kinase and ryanodine receptor (SPRY) domains. In addition, trim36 and its adjacent genes had conservative genomic synteny in different vertebrates. The upstream of the trim36 gene appeared geranylgeranyl transferase type-1 subunit beta (pggt1b), coiled-coil domain-containing protein 112 (ccdc112) and Fem-1 homolog C (fem1c) in sequence. According to the alignment result of TRIM 36 amino acid sequence of different species, the TRIM36 protein sequence of Cynoglossus semilaevis presented the highest consistency with that of Larimichthys crocea, and the homology was 89%, followed by Stegastes partitus and Oryzias latipes (88%). Moreover, the consistency between Gorilla gorilla and Macaca mulatta was the lowest of 63%. The results of RT-PCR and qRT- PCR showed that trim36 mainly expressed in ovaries and brains, which was extremely higher than that of other tissues (P＜0.05). The results of gonad in different periods by RT-PCR and qRT-PCR showed that trim36 gene began to rise at 120 d, which was consistent with the time of ovarian cavity formation, and the ovary expression was significantly higher than that in testis (P＜0.05). The in situ hybridization results demonstrated that trim36 expressed in cytoplasm at stage Ⅰ and Ⅱ of oocyte. This specific expression pattern suggested that trim36 might play a role in ovarian cavity development and stageⅠand Ⅱ of oocyte formation in half-smooth tongue sole. This study provides clues and references to support further research on trim36 gene function and TRIM family function.
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Received: 04 January 2016
Published: 20 May 2016
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