Efficient production of human acidic fibroblast growth factor in pea (Pisum sativum L.) plants by agroinfection of germinated seeds

BMC Biotechnology

Research Article

Meiying Yang¹  Jingying Liu²  Wei Li²  Xiaojuan Zhu²  Duo Xu²  Xingzhi Wang²  Yajun Fan³  Junjie Wang⁴ 

[1] College of Life Sciences, Jilin Agricultural University, 130118, Changchun, China;Institute of Genetics and Cytology, Northeast Normal University, 130024, Changchun, China;Institute of Genetics and Cytology, Northeast Normal University, 130024, Changchun, China;Department of Biology, Changchun Normal University, 130032, Changchun, China;Yunnan-Guizhou Plateau Institute of Biodiversity, Qujing Normal University, 655000, Qujing, China;Institute of Genetics and Cytology, Northeast Normal University, 130024, Changchun, China;

关键词: Vascular Endothelial Growth Factor;    Green Fluorescent Protein;    Agrobacterium;    Green Fluorescent Protein Expression;    Green Fluorescent Protein Fluorescence;   

DOI  :  10.1186/1472-6750-11-45

 received in 2010-09-15, accepted in 2011-05-06,  发布年份 2011

来源: Springer

【 摘 要 】

BackgroundFor efficient and large scale production of recombinant proteins in plants transient expression by agroinfection has a number of advantages over stable transformation. Simple manipulation, rapid analysis and high expression efficiency are possible. In pea, Pisum sativum, a Virus Induced Gene Silencing System using the pea early browning virus has been converted into an efficient agroinfection system by converting the two RNA genomes of the virus into binary expression vectors for Agrobacterium transformation.ResultsBy vacuum infiltration (0.08 Mpa, 1 min) of germinating pea seeds with 2-3 cm roots with Agrobacteria carrying the binary vectors, expression of the gene for Green Fluorescent Protein as marker and the gene for the human acidic fibroblast growth factor (aFGF) was obtained in 80% of the infiltrated developing seedlings. Maximal production of the recombinant proteins was achieved 12-15 days after infiltration.ConclusionsCompared to the leaf injection method vacuum infiltration of germinated seeds is highly efficient allowing large scale production of plants transiently expressing recombinant proteins. The production cycle of plants for harvesting the recombinant protein was shortened from 30 days for leaf injection to 15 days by applying vacuum infiltration. The synthesized aFGF was purified by heparin-affinity chromatography and its mitogenic activity on NIH 3T3 cells confirmed to be similar to a commercial product.

【 授权许可】

Unknown   
© Fan et al; licensee BioMed Central Ltd. 2011. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.