The Antibody Preparation and Proteomic Studies of the Mammalian Preimplantation Embryonic Key Proteins

• Main Authors: Chih-Yu Shen, 沈志昱
• Other Authors: Rong-Huay Juang
• Format: Others
• Language: zh-TW
• Published: 2008
• Online Access: http://ndltd.ncl.edu.tw/handle/42607805465963206394

碩士 === 國立臺灣大學 === 微生物與生化學研究所 === 96 === Among the transition stages during mammalian preimplantation development, maternal to zygotic transition (MZT) is the first key check point for embryo to maintain normal cleavage through blastocyst stage. Few researchers addressed the difference of protein expression between these transitional stages; only microarray analysis for the gene expression has been reported if any. In this study, three target proteins expressed remarkably during MZT in mouse and human embryos are used as the antigen (synthetic peptides) for antibody preparation. In addition, three unknown genes from porcine embryos were expressed and also used for antibody preparation. Finally, by using proteomic approaches, the protein patterns between mouse 2-cell and 4-cell embryos were compared. Some of the antibodies induced by synthetic polypeptides were able to binding their target pepitdes. However, none could recognize the original embryonic target protein. On the other hand, we produced monoclonal antibodies against the unknown porcine proteins, and two of them were used to produce ascites. However, these antibodies cannot recognize any protein from mouse embryo. Porcine embryo was not tested. On the two-dimensional elecetrophoresis gel, 19 protein spots reflecting the difference between mouse 2-cell and 4-cell embryo were analyzed by mass spectrometry. The results dipicted the change of the protein profile for the maternal to zygotic transition.