摘要
【目的】研究半滑舌鳎(Cynoglossussemilaevis)gata5基因(csgata5)的序列特征及该基因在半滑舌鳎性腺分化、成熟过程中的表达模式。【方法】通过克隆获得csgata5编码区序列,利用荧光定量PCR(RT-qPCR)方法检测csgata5 mRNA在不同类型性腺、胚胎和性腺发育中的表达模式,借助原位杂交技术定位csgata5 mRNA表达的细胞类型。【结果】克隆获得csgata5 cDNA序列1 777 bp,包含1 167 bp的编码区(Coding sequence,CDS),262 bp的5′UTR和348 bp的3′UTR区,预测csgata5编码388个氨基酸残基(aa),分子质量为41.9 ku,等电点为8.90,在C端含有两个典型的GATA锌指结构域;基因组分析显示csgata5位于半滑舌鳎10号染色体上,包含7个外显子和6个内含子区。RT-qPCR分析显示csgata5 mRNA在2龄雄鱼和伪雄鱼精巢中表达高于卵巢;csgata5m RNA主要定位在卵母细胞、精原细胞和精子中。胚胎发育过程中,csgata5m RNA表达在1细胞期至囊胚期呈下降趋势,之后上升,至原肠胚期达到峰值,随后降低并维持较低水平至孵化出膜。在精巢发育过程中,csgata5m RNA表达水平呈升高趋势,在1龄达到检测的峰值;在卵巢发育过程中,csgata5m RNA表达趋势类似精巢,表达水平低于同期精巢。【结论】csgata5参与半滑舌鳎原肠胚期的组织器官分化,具有促进原始生殖细胞向雄性生殖细胞分化及雄性生殖细胞发育的作用。
【Objective】In order to elucidate the gene function of gata5 of Chinese tongue sole(Cynoglossus semilaevis)(csgata5), the sequence characteristics and the expression patterns of this gene in Chinese tongue sole were studied.【Methods】The coding region of csgata5 was obtained by cloning and the expression pattern of csgata5 m RNA in embryonic development and gonadal development was detected by real-time PCR(RT-qPCR). In situ hybridization was used to localize the expression of csgata5 mRNA in the gonads.【Results】The csgata5 cDNA sequence is 1 777 bp in length and contains the entire coding region of 1 167 bp, the 262 bp 5′UTR and 348 bp 3′UTR. The location of csgata5 is on chromose 10, and it includes seven exons and six introns. The predicted protein of csgata5 contains 388 amino acid residues(aa), the molecular weight is 41.9 ku, the isoelectric point is 8.90, and it has two typical GATA zinc finger binding domains at the C-terminus. Phylogenetic analysis shows that the same GATA proteins are clustered together, and the clustering pattern is independent of species.The expression levels of csgata5 mRNA are higher in the testis of two-years old male and pseudomale than in the ovary of two-years old female and it is mainly localized in the oocytes, spermatogonia and sperm. During embryonic development, the expression levels of csgata5 m RNA decrease from the cleavage stage to the blastocyst stage, peak at the gastrula stage, and then decrease and maintain at a lower level until hatching. During the development of testis, the expression levels of csgata5 m RNA tend to increase, reaching the peak of detection at the age of one-year old. In the ovary, the expression profile of csgata5 mRNA is similar to that of the testis, and the expression level is lower than that of the testis.【Conclusion】These results show that csgata5 may be involved in the differentiation of organs in the gastrula stage of Chinese tongue sole;it has the effect of promoting the differentiation of primordial germ cells into male
作者
董忠典
张宁
崔忠凯
徐文腾
陈松林
DONG Zhong-dian;ZHANG Ning;CUI Zhong-kai;XU Wen-teng;CHEN Song-lin(Fisheries College of Guangdong Ocean University, Zhanjiang 524088, China;Yellow Sea Fisheries Research Institute, Chinese Academy of Fisheries Sciences, Qingdao 266071, China;Function Laboratory for Marine Fisheries Science and Food Production Processes / Qingdao National Laboratory for Ocean Science and Technology, Qingdao 266071, China)
出处
《广东海洋大学学报》
CAS
2019年第3期6-14,共9页
Journal of Guangdong Ocean University
基金
黄海水产研究所科研事业单位基本科研业务费项目(20603022017003)
山东省泰山学者攀登计划项目
国家自然科学基金青年科学基金项目(31402293)
广东海洋大学博士启动项目
