Molecular Cloning, Sequence Identification And Gene Expression Analysis Of Japanese Eel Dmrt2 Gene

Dmrt gene family Dmrt (double-sex and mab3-relatated transcription factor)gene is a gene family that is sexually related to gonadal development and plays an important role in the development and functional maintenance of certain organs.Among them, Dmrt2 gene is mainly involved in the development of embryos and gametes. Thus, the Dmrt gene family is important for studying sex determination and differentiation of gene families. The results of this experiment show that the Dmrt2 protein of Japanese eel contains only one functional domain, the DM domain. The molecular weight of the Dmrt2 protein is 46.82kD, the molecular formula is C2037H3281N595O617S26, the total number of atoms is 6556. The Dmrt2 protein molecule consists of Ser (S), 11.1%; Leu (L), 10.8%; Lys (K) 7.5%; and Trp (W), 0.7%. The model software predicts that the secondary structure of the Dmrt2 protein have 14?-helix, 17?-folded and has multiple rings. By comparing the domain of the Dmrt2 protein of different species by software, it was found that it was relatively conserved.The 5 'and 3' sequences of the Dmrt2 gene were analyzed by race technique to obtain the full length of the cDNA of Dmrt2 to be 1499bp. The prediction of Dmrt2 shows that there is a high homology between Japanese eel and zebrafish and Nile tilapia. In order to research the function of Dmit2 and the role of it in the differentiation of Japanese eel, the expression of genes in different tissues was analyzed by RT-PCR.The expression of Dmrt2 in the gonad was confirmed by FISH analysis. In this study,we identified the histological observation of the gonadal development of the eel and the expression of the Dmit2 gene to detect the difference in the expression of the Dmrt2 gene in the male and female individuals of the eel. The results showed that the Dmrt2 gene was expressed in a variety of tissues and was relatively high in the ovary and weakly expressed in the testis, muscle and liver tissues. For the Japanese eel treated with E2 (estradiol-17?), the Dmrt2 gene was highly expressed in the ovaries and liver, and was weakly expressed in the brain and muscle tissues. FISH results showed that the hybridization signal of Dmrt2 gene was only visible in ovary. These data will serve as a theoretical basis for further studying the sex differentiation of eel.