摘要
在先前的体外实验中，我们利用半乳糖基化组蛋白与带荧光素酶报告基因的ＥＢ病毒复制子载体ｐＥＢｌｕｃ，在体外组建了一种核酸蛋白质复合物。它可被肝细胞表面特异存在的脱唾液酸糖蛋白（Ａｓｉａｌｏｇｌｙｃｏｐｒｏｔｅｉｎ，ＡＳＧＰ）受体识别，并通过该受体介导的内吞作用将外源基因特异性导入培养的肝细胞素中表达。在此基础上，我们对该核酸蛋白复合物进行的电子显微镜观察表明，其大小在１００ｎｍ以内。在动物体内试验中证实了通过体液循环将此复合物靶向传递到肝组织的可能性。
The results we had obtained in the in vitro tests showed that pEBluc/galactosylated histone complex could be recognized and internalized via the unique asialoglycoprotein receptor on the surface of the hepatocyte. In this paper, we reported that this DNA complex could be found in a form of the doughnuts with the diameter of<100nm under electron microscope. Based on the above results, we delivered 500μg of pEBluc DNA in vivo by a 5～8 minutes perfusion into the rat peripheral circulation in the form of the pEBluc/galactosyated histone complex or plasmid only as controls, and also tested the transient luciferase activity in liver, heart, spleen, lung and kidney tissue extracts 24h later. The results demonstrated that significant level of luciferase activity could only be detected in the liver extracts of the animals received the pEBluc/galactosylated histone complex, in the themeantime, no detectable luciferase activity was noted in other tissues and controls. Dot blot analysis of total cellular DNA 24 after gene transfer showed the complete pEBluc existence and some plasmids were episomal in form of extrachromosomal DNA.
出处
《病毒学报》
CAS
CSCD
北大核心
1997年第4期309-313,共5页
Chinese Journal of Virology
基金
国家863计划基因治疗最大关键技术项目
国家高技术攀登计划
关键词
EB病毒
疱疹病毒
复制子
受体
直组织
表达
Receptor-mediated gene transfer, Electron microscope, In vivo gene transfer
