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| Transient Expression of Exogenous Gene into Plant Cell Mediated by PEI Nanovector |
| LI Ying, CUI Hai-xin, SONG Yu, LI Yao and HUANG Jin-li |
| Institute of Agricultural Environment and Sustainable Development, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China |
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摘要 This study was carried out to investigate the transfection effect of exogenous gene into plant protoplast cell mediated by polyethylenimine (PEI) nanovector, based on PEI gene delivery system in the field of medical science. PEI/DNA complexes were prepared by using PEI polymer to bind the plant expression plasmid, pCMl205-GFPn. The ability of PEI combining and protecting DNA was investigated by agarose gel electrophoresis retardation assay. The surface characteristics of PEI/DNA complexes were observed with transmission electron microscope. The transfection efficiency of Arabidopsis thaliana protoplasts mediated by PEI/DNA complexes at different N/P ratios was analyzed based on observation of transient expression of green fluorescent protein with confocal laser scanning microscope. PEI could bind and condense DNA, and form stable 100-200 nm PEI/DNA complexes when the proportion of PEl and DNA is in the range of 5:1-1:4. Transfection efficiency of PEI/DNA complexes increased with N/P ratios in range of N/P<5 and reached the highest at N/P=5, and began to decrease beyond N/P>5 as higher toxicity to cells. The transfection efficiency of PEI/DNA complexes at N/P=5 was higher than PEG. This study confirmed that PEI nanovector could effectively mediate foreign gene entering into A. thaliana protoplast cell to obtain transient expression, which may be developed as a hopeful and novel transgenic method combined with plant protoplast regeneration.
Abstract This study was carried out to investigate the transfection effect of exogenous gene into plant protoplast cell mediated by polyethylenimine (PEI) nanovector, based on PEI gene delivery system in the field of medical science. PEI/DNA complexes were prepared by using PEI polymer to bind the plant expression plasmid, pCMl205-GFPn. The ability of PEI combining and protecting DNA was investigated by agarose gel electrophoresis retardation assay. The surface characteristics of PEI/DNA complexes were observed with transmission electron microscope. The transfection efficiency of Arabidopsis thaliana protoplasts mediated by PEI/DNA complexes at different N/P ratios was analyzed based on observation of transient expression of green fluorescent protein with confocal laser scanning microscope. PEI could bind and condense DNA, and form stable 100-200 nm PEI/DNA complexes when the proportion of PEl and DNA is in the range of 5:1-1:4. Transfection efficiency of PEI/DNA complexes increased with N/P ratios in range of N/P<5 and reached the highest at N/P=5, and began to decrease beyond N/P>5 as higher toxicity to cells. The transfection efficiency of PEI/DNA complexes at N/P=5 was higher than PEG. This study confirmed that PEI nanovector could effectively mediate foreign gene entering into A. thaliana protoplast cell to obtain transient expression, which may be developed as a hopeful and novel transgenic method combined with plant protoplast regeneration.
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Received: 10 June 2011
Online: 10 June 2011
Accepted:
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Corresponding Authors:
CUI Hai-xin
E-mail: liyinglucky2001@yahoo.com.cn; zhongzhengc@yahoo.com.cn
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| About author: LI Ying, Ph D, Tel: +86-10-82105997, E-mail: liyinglucky2001@yahoo.com.cn; Correspondence CUI Hai-xin, Professor, Tel: +86-10-82106013, E-mail: zhongzhengc@yahoo.com.cn |
Cite this article:
LI Ying, CUI Hai-xin, SONG Yu, LI Yao, HUANG Jin-li.
2011.
Transient Expression of Exogenous Gene into Plant Cell Mediated by PEI Nanovector. Journal of Integrative Agriculture, 10(6): 820-826.
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