Mx—cre转基因小鼠品系的建立及其培育

将携带MX启动子调控的Cre基因真核表达载体pMx-cre线性化后,通过受卵显微注射途径制备转基因小鼠,共注射99个卵,产佴0只,利用PCR对小鼠进行筛选,以基因组Southern blot确证,最后得到一个阳性的小鼠品系,进而将其保护和扩大繁殖。     

精子为载体的基因转移研究

采用共培养和脂质体介导两种方法将金黄地鼠附睾内的精子与NFκB-luc+质粒共孵育后,再与成熟的卵母细胞进行体外受精,用PCR、Southern blot及荧光原位杂交(FISH)等技术检测外源基因在精子细胞和在受精卵中的存在及存在部位,并对相关数据进行统计学比较分析。结果表明,金黄地鼠附睾内的活精子具有主动捕获外源基因的能力,而且外源基因只存在于精子细胞的头部,而且通过受精将外源基因带入了卵细胞,脂质体的存在增加了外源基因转染精子的效率和转基因效率。Abstract: Incubate golden hamster spermatozoa and NFκB-luc+ in co-culture and liposome mediated methods, and then process the IVF with oocytes, And two groups of samples were detected by PCR, Southern Blot and FISH. Analyzed the relative data by statistics. The results showed that spermatozoa of Golden hamster epididymis can take the foreign DNA voluntarily, and the foreign DNA exist in the head of the spermatozoa can enter into zygotes by IVF. The exist of liposome can improve the foreign gene transfect ratio and transgenic ratio.