title: Identification, characterization and functional analysis of regulatory region of nanos gene from half-smooth tongue sole (Cynoglossus semilaevis).
abstract: The nanos gene encodes an RNA-binding zinc finger protein, which is required in the development and maintenance of germ cells. However, there is very limited information about nanos in flatfish, which impedes its application in fish breeding. In this study, we report the molecular cloning, characterization and functional analysis of the 3'-untranslated region of the nanos gene (Csnanos) from half-smooth tongue sole (Cynoglossus semilaevis), which is an economically important flatfish in China. The 1233-bp cDNA sequence, 1709-bp genomic sequence and flanking sequences (2.8-kb 5'- and 1.6-kb 3'-flanking regions) of Csnanos were cloned and characterized. Sequence analysis revealed that CsNanos shares low homology with Nanos in other species, but the zinc finger domain of CsNanos is highly similar. Phylogenetic analysis indicated that CsNanos belongs to the Nanos2 subfamily. Csnanos expression was widely detected in various tissues, but the expression level was higher in testis and ovary. During early development and sex differentiation, Csnanos expression exhibited a clear sexually dimorphic pattern, suggesting its different roles in the migration and differentiation of primordial germ cells (PGCs). Higher expression levels of Csnanos mRNA in normal females and males than in neomales indicated that the nanos gene may play key roles in maintaining the differentiation of gonad. Moreover, medaka PGCs were successfully labeled by the microinjection of synthesized mRNA consisting of green fluorescence protein and the 3'-untranslated region of Csnanos. These findings provide new insights into nanos gene expression and function, and lay the foundation for further study of PGC development and applications in tongue sole breeding.
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China; College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
authors.forename: Jinqiang
authors.initials: J
authors.lastname: Huang
authors.affiliation: College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China.
authors.forename: Yongjuan
authors.initials: Y
authors.lastname: Li
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
authors.forename: Changwei
authors.initials: C
authors.lastname: Shao
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China.
authors.forename: Na
authors.initials: N
authors.lastname: Wang
authors.affiliation: Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China; Laboratory for Marine Fisheries Science and Food Production Processes, Qingdao National Laboratory for Marine Science and Technology, Qingdao 266237, China. Electronic address: chensl@ysfri.ac.cn.
authors.forename: Songlin
authors.initials: S
authors.lastname: Chen
mesh_terms: D020413:3' Untranslated Regions; D000818:Animals; D005260:Female; D029941:Fish Proteins / Q000235:genetics* / Q000378:metabolism; D005399:Fishes / Q000235:genetics / Q000254:growth & development; D006066:Gonads / Q000378:metabolism; D008297:Male; D016601:RNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D012733:Sex Differentiation
chemical_list: D020413:3' Untranslated Regions; D029941:Fish Proteins; D016601:RNA-Binding Proteins
keywords: Dimorphic expression; Nanos; Primordial germ cells; Tongue sole; Visualization