Study On The Gonadal Protein And Physicochemical Properties Of Hyriopsis Schlegeli

Developmental and reproductive biology of bivalve are researched deeply all over the world. Up to now, many studies are mainly focus on dealing with the human and biomodel organism, and a number of critical genes involved in sex differentiation and reproduction have been identified. But a little available and valuable sex differentiation and reproduction information is shown in Hyriopsis schlegeli. As a kind of representative species of freshwater pearl mussels, Hyriopsis schlegeli have huge commercial values. The results not only enrich basic knowledge of developmental biology and cytological information, but also have important referenced significance in regard to artificial breed, conservation of germplasm resources and breeding of new variety of freshwater pearl mussel.A series of researches were conducted on the reproduction of Hyriopsis schlegeli, including the structure and protein components of the genital system, protein concentration and components, as well as their variation during different developmental stages, purification and partial characterization of protein, using paraffin slice, SDS-PAGE, Native-PAGE,gel filtration and Protein dialysis etc. The present research will provide foundation for further investigations on reproductive biology of Hyriopsis schlegelii in the future.The assessment can be briefly summarized as follows:1) The gonadal protein research of Hyriopsis SchlegeliIn order to observing the changes of gonad histology and gonad protein and analyzing the relationship between gonad develop and protein changes of five years old Hyriopsis Schlegeli,the methods of Paraffin sections and SDS-PAGE electrophoresis were taken respectively. The results shows:there are significant changes of gonad between breeding and non-breeding season, further more in female Hyriopsis Schlegeli, the level of gonad protein shows significant changes durning the different periods of development. From the non-breeding season to breeding season, the expression level of female gonadal protein gradually increase and reached a peak durning the breeding season, followed by a period of expression reduce and maintain a certain level. While the expression level of male gonadal proteins have shown a cycle changes, it is the highest level in the breeding season, otherwise it reached lowest level in the non-breeding season.2) Gonadal Protein and Physicochemical Properties of Hyriopsis schlegeliWe used the method of protein Dialysis to abstract the gonadal protein from mature male Hyriopsis Schlegeli, and preliminary analyze their character of chemistry and physics. The results of Native-PAGE and SDS-PAGE showed that the relative molecular weight of three protein band is 16.02KDa,14.96 KDa and 13.41 KDa respectively; there is no influence on the protein band whether existβ-mercaptoethanol or not in the buffer, so we speculate they are different isolate protein. the Native-PAGE was used to the sample after purifying. The gel which is after electrophoresis were used Sudan black B for lipoprotein-specific staining and rhodamine B for P protein-specific staining, and comparing by Coomassie blue staining.the result showed that:the negative reaction appeal in these tow staining methods,which also shows that the three proteins are not protein molecules which are rich in fat, phosphorus.