Studies Of Genes Expression Of Bovine Preimplantation Embryos Development

In order to establish the gene expression pattern of bovine 8-cell and blastocyst stages embryos and investigate the molecular mechanism of bovine embryo development, several experiments were conducted as follows:8-cell and blastocyst stages bovine em bryos were obtained through the process of oocytes fertilized in vitro (IVF) and cultured in vitro (IVC). Selection of embryonic development related genes by using single preimplantation embryo mRNA differential display reverse transcription PCR(SPEDDRT-PCR). Eleven bands were selected by using SPEDDRT-PCR. Compared the bands sequneces with GenBank after sequencing, seven bands had high homology with CALM1, HMG2L1, IK, LYAR, RPL31, RIMS2, TRM112L respectively, which are all functional gene or controlling gene; sequences of three bands were not found in GenBank; and one of the gene's function still unknow.Owing to the differential expressed genes could be false positive, RT-PCR examination was used to detect CALM1, HMG2L1, IK, LYAR, RPL31, RIMS2, TRM112L expression in bovine embryos respectively; while real-time PCR was used to detect the differerce expressions of genes in bovine 8-cell stage embryos and blastocyst stage embryos. The results showed that no expressions of HMG2L1, IK were detected in bovine 8-cell stage embryos and blastocyst stage embryos; CALM1, LYAR, RIMS2, RPL31, TRM112L express in two stages embryos; the expression of RPL31, LYAR, TRM112L, RIMS2 in 8-cell stage embryos were 3.2,46,50and 540 times higher than in blastocyst stage embryos.