Cloning And Expression Analysis Of Dmrt1in Pelodiscus Sinensis

Pelodiscus sinensis belong to the Reptilia, Testudines, Trionychidae, Pelodiscus, is atraditional precious aquatic products. The male grow fast, and having generousness skirt thanthat of females, which has higher economic value and nutritional value. The study found thatP. sinensis is no different from the sex chromosomes, the sex determination is temperaturedependent. At present, the P. sinensis gonadal differentiation mechanism is not clear, and theresearch on sex determining gene Dmrt1has not been reported yet. It is one of the membersin DMRT gene family, which is the most conservative gene of sex-determining anddifferentiation gene currently known.In order to study the influence of Dmrt1gene in P. sinensis sexual differentiation, thefull-length of Dmrt1cDNA sequence was obtained by the method of RACE, and theexpression of the gene was analyzed by Real-time PCR in different ages and different tissuesThe total length of Dmrt1gene cloned in P. sinensis is2335bp which contains164bp5′untranslated regions (UTR) and1064bp3′-UTR, and1107bp open reading frame (ORF)encoding368amino acids. The predicted molecular mass of DMRT1protein was39.20kDa,and the theoretical isoelectric point was7.58. The homology analysis showed that there were66%-97%similarities in amino acid sequence with some of previously reported other species.Real-time PCR revealed that the expression of the Dmrt1mRNA in testis of three-year-oldturtles was significantly higher than new hatched turtles (P<0.05), but there were nosignificant difference in relative expression level with one-year-old and two-year-old turtles(P>0.05). The mRNA of Dmrt1gene in ovary of new hatched and one-year-old turtles wereweakly expressed and the expression level was no significant difference (P<0.05). ThemRNA of Dmrt1gene was not expressed in ovary of two and three years old turtles, andthere were not expressed in muscle and liver of each ages. The effect of temperature on thegonad of new hatched turtles was not distinct. The results suggest that the Dmrt1gene mayplay an important role in sexual differentiation and function maintain of P. sinensis.