Studies On Developmental Gene Expression Of Goat Preimplantation Embryos

In order to establish the pattern of developmental gene expression and to understand the molecular mechanism of development block about goat preimplantation embryos, three experiments were conducted as follows:1. Establishment of silver staining mRNA differential display using single preimplantation freeze-embryo.The mice were employed in the study and its 4-cell and 8-cell stage embryos were obtained using the way of superovulation. The examination about the products of RT-PCR from single 4-cell fresh embryo and freeze embryo verified that these two kinds of the embryos had the same results. Silver staining method, in stead of radioisotope, was used in the study to display the specific bands. This newly method was useful and can avoid the radioactive pollution. The simplified one-step method and boiling method were used to recover the specific bands from silver staining PAGE. After the process of reamplifying, recovering, cloning and enzyme digesting , it was found that these two improved methods were effective and convenient when they were compared with traditional ethanol-glycogen method. One specific band in mouse 8-cell stage was selected for the further study. The sequencing and alignment result suggested that this band displayed high homology (100%) to the mouse RP23-20A6 mRNA clone. The results of this study indicated that the method of single preimplantation freeze-embryo silver staining mRNA differential display was credible and practical, and it can be used for the study of gene expression about early embryonic development.2. Studies on the developmental gene expression of preimplantation goat embryos produced in vitro. 2-cell, 4-cell and 8~16-cell stage goat embryos were obtained through the process of oocytes matured in vitro (IVM), fertilized in vitro (IVF) and cultured in vitro (IVC). Single preimplantation freeze-embryo silver staining mRNA differential display was used in the study and 16 different stage specific bands in total were screened. The sequencing and alignment results suggested that 4 differential expression bands were...