Anti-mullerian Hormone Of The Half-smooth Tongue-sole : Cloning And DKKL-1 Expression

In recent years, with the continuous development of seawater aquaculture in China, the fishery workers dedicated to research about the varieties of the high value and high nutritional value, especially in the new research focuses on halibut classes in north China. Cynoglossus semilaevis has the characteristics of growth speed, high feeding gentle, dressing percentage, sweet-meated and high market value, and that currently found one of the largest fish between the male and female growth differences. In addition, cynoglossus semilaevis is a kind of unisexual fish with the natural reverse, and ZW gender types. Fish sex related genes and molecular mechanism of sex determination are in huge demand of the fish farming industry, and it is in urgent need to overcome important scientific issues on the current international. Cynoglossus semilaevis has these features make it as pattern fish of studies of sex det ermination mechanism and gender differentiation regulation mechanism gender differ entiation regulation mechanism.According to partial sequence information obtained from the whole-genome sequencing, the study mainly do the full-length c DNA cloning, expression analysis and chromosomal localization for the cynoglossus semilaevis DKKL-1 gene. The 1508 bp DKKL-1 c DNA was obtained after RT-PCR and RACE, the complete c DNA sequence contains a 3’UTR 340 bp, 5’UTR 493 bp. The ORF encodes a putative protein with 253 amino acid residues, with a predicted molecular weight of 27.4k D and an isoelectric point(p I) of 7.98.Phylogenetic analysis showed that cynoglossus semilaevis and Haplochromis nyererei, Haplochromis boops, Maylandia zebra the highest similarity of 81%. Sequence analysis showed that cynoglossus semilaevis DKKL-1 protein has Dickkopf-N characteristic conserved family members by the first 76 to 125 amino acids, and with a signal peptide of 24 amino acids.Real-time quantitative PCR(real-time PCR) method detected that cynoglossus semilaevis DKKL-1 gene expression in the kidney and gonad tissues, small or micro expression in other tissues; DKKL-1 gene expression highest after incubation 80 d testis, indicated that it may play a role in cynoglossus semilaevis sperm cell differentiation.In short, cynoglossus semilaevis as an important breeding commercial fish, and the cloning and expression analysis of the DKKL-1 gene provides the basis for the material for the sex control in the future, and playing an important role in promoting the development of cynoglossus semilaevis aquaculture industry, to accelerate the marine fish farming pace of structural adjustment and promote sustainable development of marine fish farming.