Zhi Fei, Jiang Dong-Neng, Mustapha Umar Farouk, Li Shao-Xiang, Shi Hong-Juan, Li Guang-Li, Zhu Chun-Hua
Guangdong Provincial Key Laboratory of Aquatic Animal Disease Control and Healthy Culture, Guangdong Province Famous Fish Reproduction and Breeding Engineering Technology Research Center, Fisheries College of Guangdong Ocean University, Zhanjiang, China.
Front Genet. 2022 Aug 11;13:964150. doi: 10.3389/fgene.2022.964150. eCollection 2022.
is an isoform of the eukaryotic translation elongation factor 1 A (eEF1A) and is vital for fish ovarian development. Spotted scat () is a popular marine cultured fish species in Southern Asia and China, and its artificial reproduction is complicated, with a relatively low success ratio in practice. In this study, the gene was cloned from spotted scat. Tissue distribution analysis showed that was mainly expressed in the ovary. qRT-PCR showed that expression levels gradually decreased insignificantly in the ovaries from phase II to IV. Western blot analysis showed that was highly expressed in the ovary, while it was almost undetectable in the testis. Immunohistochemistry analysis stained mainly in the cytoplasm of the previtellogenic oocytes in ovaries of normal XX-female and sex-reversed XY-female. Aside from fish and amphibians, was also identified in some reptile species using genomic database searching. Analyses of the transcriptome data from four different fish species (Hainan medaka (), silver sillago (), Nile tilapia (), and Hong Kong catfish ()) revealed ovaries biased expression of in all, similar to spotted scat. While the neighbor genes of did not show ovary biased expression in the fish species analyzed. Bisulfite Sequencing PCR (BSP) results showed that the DNA methylation level of promoter was low in ovaries, testes, and muscles. The luciferase reporter assay demonstrated that activated expression in the presence of or . These results suggest that may be involved in regulating the early phase oocytes development of spotted scat.
是真核生物翻译延伸因子1A（eEF1A）的一种亚型，对鱼类卵巢发育至关重要。点带石斑鱼（）是南亚和中国一种受欢迎的海水养殖鱼类，其人工繁殖过程复杂，实际成功率相对较低。在本研究中，从点带石斑鱼中克隆了基因。组织分布分析表明主要在卵巢中表达。qRT-PCR显示，从II期到IV期，卵巢中的表达水平逐渐无显著下降。蛋白质免疫印迹分析表明在卵巢中高表达，而在睾丸中几乎检测不到。免疫组织化学分析显示，在正常XX雌性和性反转XY雌性卵巢中，主要在前卵黄生成期卵母细胞的细胞质中染色。除了鱼类和两栖动物，通过基因组数据库搜索在一些爬行动物种中也鉴定到了。对四种不同鱼类（海南青鳉（）、银鲈（）、尼罗罗非鱼（）和香港塘鲺（））的转录组数据进行分析，发现所有鱼类卵巢中均有偏向性表达，与点带石斑鱼类似。而在所分析的鱼类物种中，的相邻基因未显示卵巢偏向性表达。亚硫酸氢盐测序PCR（BSP）结果表明，启动子在卵巢、睾丸和肌肉中的DNA甲基化水平较低。荧光素酶报告基因检测表明，在存在或的情况下激活了表达。这些结果表明可能参与调控点带石斑鱼早期卵母细胞的发育。
