ApoCⅢ转基因小型猪的分子水平鉴定

Identification of transgenic miniature pigs express human ApoCⅢ

分析鉴定ApoCⅢ转基因阳性小型猪.从仔猪尾中提取基因组DNA,通过PCR和Southern blotting等手段进行鉴定;从仔猪肝、小肠、心、肾、肺、脾、胃以及皮肤等冰冻组织以Trizol法提取RNA,以RT-PCR手段进行检测分析.经过对18头仔猪进行PCR分析,显示其中的15头含有人ApoCⅢ基因,随后的测序和Southern blotting亦证实,猪的基因组中已经整合有人ApoCⅢ基因.RT-PCR证实人apoCⅢ基因已经在转基因小型猪的肝脏和小肠中进行了翻译.本研究成功制备了人ApoCⅢ转基因小型猪,可作为高脂血症与动脉粥样硬化疾病之间关系很有价值的研究模型.

For identification of positive ApoCⅢ transgenic miniature pigs, genomic DNA was extracted from the umbilical cord for PCR and Southern blotting. To examine the expression of human ApoCⅢ in transgenic pig tissue, total RNA was isolated from different tissues for RT-PCR. Among the 18 natural-born piglets, 15 were confirmed as human ApoCⅢ transgenic pigs by PCR genotyping, South- ern blot analysis also confirmed the integration of human ApoCⅢ fragments into the transgenic pigs. By the use of RT-PCR, we found that transgenic human ApoCⅢ was exclusively expressed in the liver and intestine in the transgenic miniature pigs. We have successfully generated a novel hypertriglyceri- demic ApoCⅢ transgenic miniature pig model that could be of great value for studies on hyperlipi- demia in relation to atherosclerotic disorders.