精子干细胞转染法制备转基因兔的研究

Research on Transfection Spermatogenous Cells in the Production of Transgenic Rabbits

采用直接注射的方法 ,将脂质体包裹的含人乳铁蛋白基因重组质粒 pLNCXHLF注入兔睾丸组织中 ,一个月后与正常雌兔交配。所产仔兔经PCR、Southern检测证明获得了转基因兔 ,转基因阳性率平均为 35 %。实验结果表明 ,通过注射可将外源基因导入到曲细精管中 ,进而完成对精子干细胞的转染 ,获得携带外源基因的成熟精子 ,受精后可得到转基因动物。该方法是一种简捷、有效的新途径 ,既节省时间又降低了成本 ,为利用精子载体制备转基因动物的研究提供了很有价值的参考。

The recombinant plasmids pLNCXHLF constructed by eukaryotic expression vectors pLNCX fused with human lactoferritin (hLF) gene were injected into testes of a healthy male rabbit directly using liposome entrapment.One month later, the transfected male rabbit was fertilized with female rabbits. The analysis of PCR and Southern hybridization of newborn rabbits genomic DNA shows that transgenic rabbits were produced and the positive rate is 35%. The results of this experiment shows that the transgenic animals can be introduced by gene transfering into spermatogenous cells by exogenous gene injection into seminiferous tubules then get a mature sperm to fertilize. The system, saving time and lowering costs, provides production transgenic animals with a reference result.