title: Efficient production of human acidic fibroblast growth factor in pea (Pisum sativum L.) plants by agroinfection of germinated seeds.
abstract: BACKGROUND
  For 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.
  
  
  RESULTS
  By 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.
  
  
  CONCLUSIONS
  Compared 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.
authors.affiliation: Institute of Genetics and Cytology, Northeast Normal University, Changchun 130024, China.
authors.forename: Yajun
authors.initials: Y
authors.lastname: Fan
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.forename: Junjie
authors.initials: J
authors.lastname: Wang
authors.forename: Jingying
authors.initials: J
authors.lastname: Liu
authors.forename: Meiying
authors.initials: M
authors.lastname: Yang
authors.forename: Duo
authors.initials: D
authors.lastname: Xu
authors.forename: Xiaojuan
authors.initials: X
authors.lastname: Zhu
authors.forename: Xingzhi
authors.initials: X
authors.lastname: Wang
mesh_terms: D000818:Animals; D016220:Fibroblast Growth Factor 1 / Q000235:genetics* / Q000378:metabolism; D020868:Gene Silencing; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D051379:Mice; D041681:NIH 3T3 Cells; D018532:Pisum sativum / Q000235:genetics*; D018515:Plant Leaves / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics* / Q000378:metabolism; D012231:Rhizobium / Q000235:genetics; D012639:Seeds / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D016220:Fibroblast Growth Factor 1; D049452:Green Fluorescent Proteins