我们用50000猪卵母细胞免疫大鼠，建立了抗猪卵母细胞特异单克隆抗体库。筛选单克隆抗体库，我们得到了两个单克隆抗体- - BM26和BM199,它们分别识别一个32KD的BM26蛋白和一个卵母细胞特异表达的蛋白BM199。初步研究结果表明BM26出现在刚刚恢复减数分裂的卵母细胞的细胞质，随着卵母细胞进入减数分裂，BM26进入卵母细胞的细胞核，如用抗体阻断BM26蛋白质进入细胞核，则阻断卵母细胞减数分裂 在GVBD，BM26蛋白也存在于受精卵的原核；在3T3细胞中，BM26基因的表达与细胞周期紧密相关联。卵母细胞特异蛋白BM199的表达和表达水平与卵母细胞生长紧密相关。我们的初步研究结果表明Bm26和BM199是有待于研究的新功能蛋白质。本课题拟定克隆BM26和BM199基因，用抑制或提高基因在卵母细胞中表达的方法，来研究BM26和BM199在卵母细胞生长、成熟和受精后基因表达的功能。

In order to find novel proteins in pig oocytes, we immunized rats with 50000 pig oocytes. An anti- pig oocyte monoclonal antibody library was made with the spleen cells obtained from the immunized rats. Total of 500 monoclonal antibodies were produced. Western blots with total pig oocyte proteins were used to screen the library. Two antibodies were found to be the most interesting. The first one is BM26 monoclonal antibody which picked up a 32kD protein on western blot. Immunostaining of oocytes showed that BM26 proteins were present in cytoplasm of the oocytes just released from follicles. As the maturation progressed, BM26 proteins relocated into the nuclear of GV2 stage oocytes. The maturation of oocytes stopped at GVDB stage when the relocation of BM26 proteins in the GV oocytes was blocked by antibody injection. In 3T3 cells the expression of BM26 proteins were closely related to cell cycle and were detected at G2 and M phase. The BM26 proteins were located in the nuclear in the G2 cells and were overlapped with condensed chromatins and spindles in M phase cells. The second one is BM199 monoclonal antibody that recognized oocyte specific proteins which were detected in primordial oocytes. With progression of oocyte growth, the expression levels of BM199 proteins increased. As shown by western blot, BM199 antibody picked up a 50kD protein in oocytes just released from follicles. But in MII oocytes it not only detected the 50kD protein, but also a 70kD protein. In this research proposal we propose that we will first clone the BM26 and BM199 genes. We will then use up and down regulation approaches to study the functions of BM26 and BM199 proteins during oocyte maturation and their roles in the regulation of gene expression after fertilization.