钠米颗粒介导质粒DNA转染体外真核细胞  

Transfection of Eukaryotic Cells in vitro Using Chitosan-16/Plasmid Complex

作　　者：金华利[1] 张富春[1] 张爱莲[1] 李轶杰[1] 王宾[1] 

机构地区：[1]新疆大学生命科学与技术学院新疆生物资源基因工程重点实验室,乌鲁木齐830046

出　　处：《中国生物工程杂志》2003年第10期71-75,80,共6页China Biotechnology

基　　金：国家 8 63计划 (2 0 0 1AA2 13 111);国家自然科学基金资助项目(3 0 2 712 2 0 )

摘　　要：DNA传递是基因表达与功能研究及其医学应用的重要技术 ,安全高效的DNA传递一直是研究者期待的目标。利用一种新的阳离子多聚物脱乙酰甲壳胺 1 6介导重组质粒pcDNA3 vp1转染COS 7细胞 ,RT PCR可检测到目的基因vp1在mRNA水平的表达 ,实时定量PCR结果表明其转染效率介于脂质体与磷酸钙法之间 ,同时还对转染条件进行了探讨。DNA结合分析发现脱乙酰甲壳胺 1 6能够与DNA形成核酸纳米颗粒 ,提高DNA稳定性 ,促进真核细胞转染效率的提高。这些结果表明脱乙酰甲壳胺 1 6确能做为一种新型的非病毒纳米DNA传递载体 。DNA delivery is an important technology in the study of expression and function of genes. Many laboratories are making great efforts to find a safe and high efficient delivery vehicle. The construct of pcDNA3/vp1 was transfected into eukaryotic cells in vitro with low molecular weigh of chitosan, a cational polymer designated as chitosan 16. After expressions of HPRT and vp1, the efficiencies of such facilitative transfection were determined by RT PCR and real time quantitative PCR technologies compared with transfected using the lipofectin and calcium phosphate methods. The results showed that higher expression of the vp1 at mRNA level mediated with the chitosan 16/DNA was observed in COS 7 than that from the calcium phosphate and lipfectin. Formation of nano particles of chitosan 16 with DNA was revealed under electron microscope and this particle complex may improve the stability of DNA and the efficiency of transfection. It could be applicable to non viral based delivery system for gene therapy as well as DNA vaccination applications.

关 键 词：脱乙酰甲壳胺 DNA传递 实时定量PCR 核酸纳米颗粒 

分 类 号：Q75[生物学—分子生物学] Q78