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

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 of PEI/DNA at different charge ratios (N/P) was analyzed and compared with PEG. 【Result】 When the quality rate of PEI and DNA is in range of 5﹕1-1﹕4, PEI could bind and condense DNA, and form stable PEI/DNA complexes with nanoparticles diameter in 100-200 nm. Transformation efficiency of PEI/DNA can be improved with the increase of N/P ratios and reached highest at N/P=5. There was higher transformation efficiency of PEI/DNA complexes than PEG at N/P=5, but began to decrease at N/P＞5 with higher toxicity to cells. 【Conclusion】 The experimental results confirmed that PEI is a new kind of efficient gene vector for transferring genes into Arabidopsis thaliana protoplasts.

Key words: polyethylenimine, nano-gene vector, plant cell, transient expression, Arabidopsis thaliana