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牙鲆(Paralichthys olivaceus)sox3基因的分子克隆、启动子分析及表达谱

Molecular Cloning, Promoter Analysis and Expression Profiles of the sox3 Gene in Japanese Flounder, Paralichthys olivaceus.

作者信息

Gao Jinning, Li Peizhen, Zhang Wei, Wang Zhigang, Wang Xubo, Zhang Quanqi

机构信息

Center for Developmental Cardiology, Institute for Translational Medicine, College of Medicine, Qingdao University, 38 Dengzhou Road, Qingdao 266021, China.

Key Laboratory of Marine Genetics and Breeding (MGB), Ministry of Education, College of Marine Life Sciences, Ocean University of China, 5 Yushan Road, Qingdao 266003, China.

出版信息

Int J Mol Sci. 2015 Nov 24;16(11):27931-44. doi: 10.3390/ijms161126079.

Abstract

Sox3, which belongs to the SoxB1 subgroup, plays major roles in neural and gonadal development. In the present study, Japanese flounder Paralichthys olivaceus sox3 gene (Posox3) and its promoter sequence were isolated and characterized. The deduced PoSox3 protein contained 298 amino acids with a characteristic HMG-box domain. Alignment and phylogenetic analyses indicated that PoSox3 shares highly identical sequence with Sox3 homologues from different species. The promoter region of Posox3 has many potential transcription factor (TF) binding sites. The expression profiles of Posox3 in different developmental stages and diverse adult tissues were analyzed by quantitative real-time RT-PCR (qRT-PCR). Posox3 mRNA was maternally inherited, and maintained at a considerably high expression level between the blastula stage and the hatching stage during embryonic development. Posox3 was abundantly expressed in the adult brain and showed sexually dimorphic expression pattern. In situ hybridization (ISH) was carried out to investigate the cellular distribution of Posox3 in the ovary, and results showed the uniform distribution of Posox3 throughout the cytoplasm of oogonia and stage I-III oocytes. These results indicate that Posox3 has potentially vital roles in embryonic and neural development and may be involved in the oogenesis process. Our work provides a fundamental understanding of the structure and potential functions of Sox3 in Paralichthys olivaceus.

摘要

属于SoxB1亚组的Sox3在神经和性腺发育中起主要作用。在本研究中,分离并鉴定了牙鲆(Paralichthys olivaceus)的sox3基因(Posox3)及其启动子序列。推导的PoSox3蛋白包含298个氨基酸,具有特征性的HMG-box结构域。比对和系统发育分析表明,PoSox3与来自不同物种的Sox3同源物具有高度相同的序列。Posox3的启动子区域有许多潜在的转录因子(TF)结合位点。通过定量实时RT-PCR(qRT-PCR)分析了Posox3在不同发育阶段和不同成体组织中的表达谱。Posox3 mRNA是母系遗传的,并且在胚胎发育的囊胚期和孵化期之间保持相当高的表达水平。Posox3在成体脑中大量表达,并表现出性别二态性表达模式。进行原位杂交(ISH)以研究Posox3在卵巢中的细胞分布,结果显示Posox3在卵原细胞和I-III期卵母细胞的整个细胞质中均匀分布。这些结果表明,Posox3在胚胎和神经发育中可能具有至关重要的作用,并且可能参与卵子发生过程。我们的工作为牙鲆中Sox3的结构和潜在功能提供了基本的认识。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/eb0a/4661933/2d70ad79a516/ijms-16-26079-g001.jpg

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