title: Evaluation of somatic embryos of alfalfa for recombinant protein expression.
abstract: Somatic embryos of alfalfa can accumulate higher levels of recombinant proteins comparing to vegetative organs. Somatic embryos may be explored as a new system for new protein production for plants. Plants have been explored via genetic engineering as an inexpensive system for recombinant protein production. However, protein expression levels in vegetative tissues have been low, which limits the commercial utilization of plant expression systems. Somatic embryos resemble zygotic embryos in many aspects and may accumulate higher levels of proteins as true seed. In this study, somatic embryo of alfalfa (Medicago sativa L.) was investigated for the expression of recombinant proteins. Three heterologous genes, including the standard scientific reporter uid that codes for β-glucuronidase and two genes of interest: ctb coding for cholera toxin B subunit (CTB), and hIL-13 coding for human interleukin 13, were independently introduced into alfalfa via Agrobacterium-mediated transformation. Somatic embryos were subsequently induced from transgenic plants carrying these genes. Somatic embryos accumulated approximately twofold more recombinant proteins than vegetative organs including roots, stems, and leaves. The recombinant proteins of CTB and hIL-13 accumulated up to 0.15 and 0.18 % of total soluble protein in alfalfa somatic embryos, respectively. The recombinant proteins expressed in somatic embryos also exhibited biological activities. As somatic embryos can be induced in many plant species and their production can be scaled up via different avenues, somatic embryos may be developed as an efficient expression system for recombinant protein production.
authors.affiliation: Department of Biology, University of Western Ontario, London, ON, Canada.
authors.forename: Guohua
authors.initials: G
authors.lastname: Fu
authors.forename: Vojislava
authors.initials: V
authors.lastname: Grbic
authors.forename: Shengwu
authors.initials: S
authors.lastname: Ma
authors.forename: Lining
authors.initials: L
authors.lastname: Tian
mesh_terms: D060054:Agrobacterium / Q000235:genetics; D002772:Cholera Toxin / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D017930:Genes, Reporter; D005818:Genetic Engineering; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism*; D018793:Interleukin-13 / Q000235:genetics / Q000378:metabolism*; D000455:Medicago sativa / Q000235:genetics / Q000378:metabolism*; D058634:Molecular Farming / Q000379:methods*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D058439:Plant Somatic Embryogenesis Techniques; D018547:Plant Stems / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012639:Seeds / Q000235:genetics / Q000378:metabolism; D014170:Transformation, Genetic; D019076:Transgenes
chemical_list: D018793:Interleukin-13; D011994:Recombinant Proteins; D002772:Cholera Toxin; D005966:Glucuronidase