PEI 介导外源基因进入植物细胞的瞬时表达.pdfVIP

中国农业科学 2009,42(6):1918-1923 Scientia Agricultura Sinica doi: 10.3864/j.issn.0578-1752.2009.06.006 PEI介导外源基因进入植物细胞的瞬时表达 李 颖，崔海信，宋 瑜，李 瑶，黄金丽 （中国农业科学院农业环境与可持续发展研究所，北京 100081 ） 摘要：【目的】医学方面的大量研究证实，聚乙烯亚胺（polyethylenimine，PEI）作为一种新型阳离子多聚 物基因载体可以吸附和浓缩 DNA，通过与细胞膜亲和粘附和细胞吞噬作用运载外源基因进入细胞并实现表达。本 实验研究旨在探索 PEI 作为非病毒基因载体介导外源基因进入植物细胞并获得瞬时表达的可能性。【方法】制备 PEI/DNA 复合物，利用凝胶阻滞分析 PEI 与 DNA 的结合情况，采用电子显微镜观察 PEI/DNA 复合物的形态。以绿 色荧光蛋白基因为报告基因研究不同 N/P 比条件下 PEI/DNA 复合物对拟南芥原生质体细胞的转化效率，并与 PEG 转化方法进行比较分析。【结果】PEI与DNA 的质量比为5﹕1～1 ﹕4，PEI可以与DNA稳定结合，形成粒径约100～ 200 nm的球形复合物。PEI/DNA复合物对拟南芥原生质体细胞的转化效率随其N/P比的增大而提高，N/P＝5时PEI 的转化效率达到最高且明显高于PEG介导的转化效率，N/P＞5时转化效率反而下降，容易使细胞破碎和变形。【结 论】PEI作为一种新型的外源基因运送载体对拟南芥原生质体细胞具有良好的介导基因转移效果。 关键词：聚乙烯亚胺；纳米基因载体；植物细胞；瞬时表达；拟南芥 Transient Expression of Transferred Gene Induced by Polyethylenimine in Plant LI Ying, CUI Hai-xin, SONG Yu, LI Yao, HUANG Jin-li (Institute of Agricultural Environment and Sustainable Development, Chinese Academy of Agricultural Sciences, Beijing 100081) Abstract: 【Objective 】 The experimental research was carried out to investigate the possibility and method of gene delivery into plant cells by nano-vector polyethylenimine (PEI) and obtain transient expression of gene, based on some research results that PEI can condense DNA into granules of nanometer size for delivering gene into cells in the field of medical science. 【Method 】 PEI/DNA complexes were prepared. The ability of PEI to combine and protect DNA was investigated by agarose gel electrophoresis retardation assay. The surface characteristics of PEI/DNA complexes were observed with transmission electron microscope. For Arabidopsis thaliana protoplasts, transformation efficiency