摘要
关节腔内注射裸露ＤＮＡ，是实现关节内基因转移的一种最简便的方法，但其机制尚不了解。本实验利用差速离心法，提取类风湿性关节炎患者关节滑膜细胞的非核膜蛋白，以大鼠肝细胞非核膜蛋白作为对照，应用32Ｐ标记的 ＤＮＡ，进行 ＤＮＡ膜蛋白结合实验。我们发现在滑膜细胞微粒体膜的抽提物中存在一种非核 ＤＮＡ结合蛋白质，分子量约为８３ＫＤａ，这种蛋白质能特异地与双链线状ＤＮＡ结合，可能在关节滑膜介导的基因转移中起一定作用。
It is one of the simplest methods that gene transfer in the is achieved after intra-articular injection of naked plasmid DNA into the knee joint.But the mechanism by which synovial tissue internalizes plasmid DNA is unknown. In the present study, we extracted the non-nuclear membrane proteins from both synovial tissue of RA patients and liver tissue of rats by means of differential centrifugation.The non-nuclear mem- brane protein is served as control. 32P labeling of double-strand DNA and overlay protocol were used to identify membrane-associated DNA binding proteins.We found a 83Kda protein that bound double-strand DNA specifically and with high sanity in the synovial cell membrane ex- tracts. The membrane-associated DNA binding protein may mediated the uptake and expression of plasmid DNA by synovia cells and play a role in the the synovial mediated gene transfer.
出处
《中国运动医学杂志》
CAS
CSCD
北大核心
1999年第2期103-105,共3页
Chinese Journal of Sports Medicine
关键词
关节炎
关节腔注射
裸露质粒DNA
基因转移
non-nuclear membrane-associated DNA binding protein, gene transfer, articular synovium, naked plasmid DNA
