Proteome rebalancing in soybean seeds can be exploited to enhance foreign protein accumulation.
Seeds possess a high intrinsic capacity for protein production that makes them a desirable bioreactor platform for the manufacture of transgenic products. One strategy to enhance foreign protein production involves exchanging the capacity to produce intrinsic proteins for the capacity to produce a high level of foreign proteins. Suppression of the alpha/alpha' subunit of beta-conglycinin storage protein synthesis in soybean has been shown previously to result in an increase in the accumulation of the glycinin storage protein, some of which is sequestered as proglycinin into de novo endoplasmic reticulum (ER)-derived protein bodies. The exchange of glycinin for conglycinin is quantitative, with the remodelled soybeans possessing a normal protein content with an altered proteome. The green fluorescent protein (GFP)-kdel reporter was transferred in a construct using the glycinin promoter and terminator to mimic glycinin gene expression. When expressed in soybean seeds, GFP-kdel accreted to form ER-derived protein bodies. The introgression of GFP-kdel into the alpha/alpha' subunit of the beta-conglycinin suppression background resulted in a fourfold enhancement of GFP-kdel accumulation to > 7% (w/w) of the total protein in soybean seeds. The resulting seeds accumulated a single population of ER membrane-bound protein bodies that contained both GFP-kdel and glycinin. Thus, the collateral proteome rebalancing that occurs with the suppression of intrinsic proteins in soybean can be exploited to produce an enhanced level of foreign proteins.
Schmidt
Monica A
MA
Plant Genetics Research Unit USDA/ARS, Donald Danforth Plant Science Center, 975 N Warson Road, St Louis, MO 63132, USA.
Herman
Eliot M
EM
D052179:Antigens, Plant; D015180:Electrophoresis, Gel, Two-Dimensional; D004721:Endoplasmic Reticulum / Q000378:metabolism; D018506:Gene Expression Regulation, Plant; D017343:Genes, Plant; D017930:Genes, Reporter; D005916:Globulins / Q000235:genetics*; D049452:Green Fluorescent Proteins / Q000235:genetics; D007525:Isoelectric Focusing; D030821:Plants, Genetically Modified / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020543:Proteome / Q000235:genetics / Q000378:metabolism; D055314:Seed Storage Proteins; D012639:Seeds / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D030262:Soybean Proteins / Q000235:genetics*; D013025:Glycine max / Q000235:genetics* / Q000378:metabolism
D052179:Antigens, Plant; D005916:Globulins; D020543:Proteome; D055314:Seed Storage Proteins; D030262:Soybean Proteins; C013894:beta-conglycinin protein, Glycine max; D049452:Green Fluorescent Proteins; C011474:glycinin