重组质粒pcDNKCpG在动物体内分布、残留及水平传播检测  

biodistribution, persistence and horizontal transmission of the recombinant pcDNKCpG in animals

作　　者：张泉[1] 于可响[1] 严彩红[1] 郭晓宇[1] 孙怀昌[1] 朱鸿飞[2] 

机构地区：[1]扬州大学兽医学院,扬州225009 [2]中国农业科学院生物制品工程技术中心,北京100081

出　　处：《高技术通讯》2006年第6期629-632,共4页Chinese High Technology Letters

基　　金：863计划（2003AA243070和2005AA246020）资助项目.

摘　　要：通过肌肉注射的途径将重组质粒pcDNKCpG导入到鸡和猪体内，提取不同时间段的动物组织DNA，特异性的PCR扩增结果提示重组质粒pcDNKCpG主要分布在鸡和猪的血液和注射部位肌肉中，残留时间分别不超过16天和8天；凝胶电泳回收的组织DNA经过酶切、电泳和转膜后的Southem检测表明，重组质粒pcDNKCpG没有整合到鸡和猪的任何组织DNA中去；地高辛标记的CpG核酸探针的Southem检测结果显示重组质粒pcDNKCpG没有整合入宿主的基因组中；特异性的PCR扩增结果证实重组质粒pcDNKCpG没有转移到其他的环境微生物中去。因此，重组质粒pcDNKCpG具有良好的生物安全性。The recombinant PeDNKCpG was injected into chickens and pigs intamuscularly. After tissue DNA extraction and specific PCR amplification in different periods, the results showed that the biodistribution of plasmid was mainly located in the blood and the muscle where the plasmid was injected, and the plasmid existed for no more than 16 days and 8 days, respectively. The tissue DNA was recovered by gel electrophoresis and detected by Southern blotting after enzyme digestion, electrophoresis and transferring to a piece of membrane, and the results revealed the plasmid peDNKCpG did not integrate into the host genome. Meanwhile, the specific amplification of PCR indicated that the plamsid peDNKCpG did not integrate into any other microorganism in environment. Therefore, the recombinant peDNKCpG had a good biosafetv.

关 键 词：分布 残留 PCR Southem杂交 生物安全 

分 类 号：Q78[生物学—分子生物学]