渔业科学进展2018，Vol.39Issue(3)：126-136,11.DOI:10.19663/j.issn2095-9869.20170401001
三疣梭子蟹Drosha和Exportin 5基因克隆及其在性腺发育过程中的表达分析
Molecular Cloning of Drosha and Exportin 5 and Their Expression During Gonadal Development in the Swimming Crab (Portunus trituberculatus)
摘要
Abstract
In the present study, full-length complementary DNA (cDNA) of Drosha and Exportin 5 involved in the pathway of microRNA (miRNA) biogenesis of the swimming crab Portunus trituberculatus were cloned by rapid amplification of cDNA ends (RACEs). The length of the Drosha gene is 3443 bp, which is predicted to encode a polypeptide of 1038 amino acids, containing two ribonuclease (RNase) Ⅲ domains (RⅢDa and RⅢDb) and a double-stranded RNA binding domain (dsRBD). The full-length of Exportin 5 is 5000 bp, which is predicted to encode a polypeptide of 1208 amino acids, containing an importin-β N-terminal domain (IBN-N) and an exportin 1-like domain (XPO-1). Homology analysis revealed that Drosha in P. trituberculatus is highly similar to those in some species and shares the highest similarity with that in Marsupenaeus japonicus (94％). Quantitative real-time PCR (qRT-PCR) analysis showed that Drosha and Exportin 5 are expressed in all the tested tissues, and were highly expressed in the ovary and hepatopancreas tissues (P<0.05). At different stages of testis development, the expression of the genes Drosha and Exportin 5 shared the same trend, and both showed the highest expression at stage Ⅱ. During ovarian development, the expression level of the genes Drosha and Exportin 5 increased gradually from phase Ⅱ to phase Ⅴ. The results suggest that the genes Drosha and Exportin 5 cooperatively regulate the gonadal development of P. trituberculatus in an miRNA-dependent manner. Furthermore, the results provide useful information for studies on the regulation of gonadal development in P. trituberculatus.关键词
三疣梭子蟹/Drosha/Exportin5/基因克隆/基因表达Key words
Portunus trituberculatus/Drosha/Exportin 5/Gene cloning/Expression analysis分类
农业科技引用本文复制引用
张小辉,孟宪亮,高保全,刘萍,王竹青,张杰,蔡影..三疣梭子蟹Drosha和Exportin 5基因克隆及其在性腺发育过程中的表达分析[J].渔业科学进展,2018,39(3):126-136,11.基金项目
国家自然科学基金项目(41576147 （）
41306178)、泰山领军人才工程高效生态农业创新类计划项目(LJNY2015002)和中国水产科学研究院基本科研业务费(2018HY-ZD0201)共同资助 [This work was supported by National Natural Science Foundation of China (41576147 （LJNY2015002）
41306178), Program of Shandong Leading Talent (LJNY2015002), and Central Public-Interest Scientific Institution Basal Research Fund, CAFS(2018HY-ZD0201)]. （LJNY2015002）
