Preliminary Study Of Transgenic Mouse Model Using Sperm Vector

SMGT (Sperm-mediated gene transfer) is a method that uses sperm as the vector to introduceexogenous gene into oocyte when fertilized. SMGT is one of the most simple and efficient methods in thegene transfer of animal research, so ICR mouse were used as experiment materials to make transgenic micemodel by sperm-mediated method, and then, the feasibility, advantage and disadvantage was analyzed anddiscussed, in this paper, according to the results.The research of the mouse sperm with exogenous DNA: the efficiency of mouse sperm transfectingthe exogenous DNA was studied by using the DIG end labeled technology and immunohistochemistrytechnology. The result showed that: the efficiency of transfecting positive rate of individual mouse spermwas distinct difference, and the average number was 13%. Mouse sperms had the unprompted capability ofintegrating with exgenous DNA, and the place of integration congregate was in the postacrosomal region.The integrated exgenous DNA was stuck to the external surface of the sperm membrane firstly and thentransfered into the inside of the cell, but not all the integrated DNA was inner transfered.The study on Sperm-mediated gene transfer: The acrosomal reaction was evaluated with thetechnology of Coomassie brilliant blue stained, and the appropriate fertilization medium in vitro waselected .By Using IVF, GFP was transfected in vitro, the mature oocytes were fertilized, and then thezygotes were incubated in vitro. The eygotes were observed through the fluorescence microscopy, and theresult was 4.7%. The above results showed SMGT was an effective and feasible method.The construction of HBVBP expressed vector: the vector that was expressed by HBV was constructed byPCR and gene recombinant. The correctness of the vector sequence was testified by endonuclease digestionand the sequence analysis. This vector was applied to transfect mice sperm to make animal disease models.According to the above research, the foundation of making animal disease models and using SMGT toobtain transgenic mouse were established. But in this research HBVbp was not transfected, the furtherresearch will be needed.