Abstract
Erythropoietin (EPO) is a glycoprotein used for curing human anemia by regulating the differentiation of erythroid progenitors and the production of red blood cells. To examine the expression of recombinant EPO in plants, pPEV-EP21, in which human epo cDNA under the control of the CaMV 35S promoter, was introduced into tobacco and Arabidopsisvia Agrobacterium tumefaciens-mediated transformation. The RNA expression level of epo in the transgenic lines was initially estimated by Northern blot analysis. Two transgenic lines, which exhibited a high expression level of epo mRNA determined by Northern analysis, were chosen for Western blot analysis to examine the production of EPO proteins. Those two lines, EP21-12 and EP21-14, revealed detectable bands on the immunoblot. Interestingly, constitutive expression of the human epo gene affected the morphologies in transgenic plants such that vegetative growth of transgenic tobacco was retarded, and male sterility was induced in transgenic tobacco and Arabidopsis
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- cDNA:
-
complementary DNA
- CaMV:
-
cauliflower mosaic virus
- EPO:
-
erythropoietin
- PCR:
-
polymerase chain reaction
References
M Cabanes-Macheteau AC Fitchette-Laine C Loutelier-Bourhis C Lange ND Vine JK. Ma et al. (1999) ArticleTitleN-glycosylation of a mouse IgG expressed in transgenic tobacco plants Glycobiology 9 365–372
P Chomczynski (1992) ArticleTitleOne-hour downward alkaline capillary transfer for blotting of DNA and RNA Anal Biochem 201 134–139
SJ Clough AF Bent (1998) ArticleTitleFloral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana Plant J 16 735–743
H Daniell SJ Streatfield K Wycoff (2001) ArticleTitleMedical molecular farming: production of antibodies, biopharmaceuticals and edible vaccines in plants Trends Plant Sci 6 219–226
MS Dodal FF Wang E Goldwasser (1985) ArticleTitleThe role of carbohydrate in erythropoietin action Endocrinology 116 2293–2299
S Dorion S Lalonde HS Saini (1996) ArticleTitleInduction of male sterility in wheat by meiotic-stage water deficit is preceded by a decline in invertase activity and changes in carbohydrate metabolism in anthers Plant Physiol 111 137–145
R Ehneß T Roitsch (1997) ArticleTitleCo-ordinated induction of mRNAs for extracellular invertase and a glucose transporter in Chenopodium rubrum by cytokinins Plant J 11 539–548
S Elliott J Giffin S Suggs EP Lau AR Banks (1989) ArticleTitleSecretion of glycosylated human erythropoietin from yeast directed by the α-factor leader region Gene 79 167–180
R Fischer N Emans (2000) ArticleTitleMolecular farming of pharmaceutical proteins Transgenic Res 9 279–299
G Giddings G Allison D Brooks A Carter (2000) ArticleTitleTransgenic plants as factories for biopharmaceuticals Nat Biotechnol 18 1151–1155
M Goetz DE Godt A Guivarc’h U Kahmann D Chriqui T Roitsch (2001) ArticleTitleInduction of male sterility in plants by metabolic engineering of the carbohydrate supply Proc Natl Acad Sci USA 98 6522–6527
A Guivarc’h J Rembur M Goetz T Roitsch M Noin T. Schmüling et al. (2002) ArticleTitleLocal expression of the ipt gene in transgenic tobacco (Nicotiana tabacum L cv. SR1) axillary buds establishes a role for cytokinins in tuberization and sink formation. J Exp Bot 53 621–629
M Holsters D de Waele A Depicker E Messens M van Montagu J Schell (1978) ArticleTitleTransfection and transformation of Agrobacterium tumefaciens Mol Gen Genet 163 181–187
RB Horsch J Fry N Hoffmann J Neidermeyer SG Rogers RT Fraley (1988) Leaf disc transformation Plant Molecular Biology Manual. A5. (pp. 1–9) Kluwer Academic Publishers New York
LW Hynes J Peng DE Richards NP Harberd (2003) ArticleTitleTransgenic expression of the Arabidopsis DELLA proteins GAI and gai confers altered gibberellin response in tobacco Transgenic Res 12 707–714
K Jacobs C Shoemaker R Rudersdorf SD Neill RJ Kaufman A. Mufson et al. (1985) ArticleTitleIsolation and characterization of genomic and cDNA clones of human erythropoietin Nature 313 806–810
W Jelkmann (1992) ArticleTitleErythropoietin: Structure, control of production, and function Physiol Rev 72 449–489
CS Kim CH Lee JS Shin YS Chung NI Hyung (1997) ArticleTitleA simple and rapid method for isolation of high quality genomic DNA from fruit trees and conifers using PVP Nucleic Acids Res 25 1085–1086
KE Koch (1996) ArticleTitleCarbohydrate-modulated gene expression in plants Annu Rev Plant Physiol Plant Mol Biol 47 509–540
PH Lai R Everett FF Wang T Arakawa E Goldwasser (1986) ArticleTitleStructural characterization of human erythropoietin J Biol Chem 261 3116–3121
JW Larrick DW Thomas (2001) ArticleTitleProducing proteins in transgenic plants and animals Curr Opin Biotech 12 411–418
S Lee-Huang (1984) ArticleTitleCloning and expression of human erythropoietin cDNA in Escherichia coli Proc Natl Acad Sci USA 81 2708–2712
FK Lin S Suggs CH Lin JK Browne R Smalling JC. Egrie et al. (1985) ArticleTitleCloning and expression of the human erythropoietin gene Proc Natl Acad Sci USA 82 7580–7584
S Matsumoto K Ikura M Ueda R Sasaki (1995) ArticleTitleCharacterization of a human glycoprotein (erythropoietin) produced in cultured tobacco cells Plant Mol Biol 27 1163–1172
T Miyake CK Kung E Goldwasser (1977) ArticleTitlePurification of human erythropoietin J Biol Chem 252 5558–5564
K Oh BY Cheon SH Cho HQ Truong SH Ok JU. Jeung et al. (2003) ArticleTitleExpression of the bovine growth hormone alters the root morphology in transgenic tobacco plants Transgenic Res 12 363–367
FW Quelle LF Caslake RE Burkert DM Wojchowski (1989) ArticleTitleHigh-level expression and purification of a recombinant human erythropoietin produced using a baculovirus vector Blood 74 652–657
T Roitsch R Ehneß (2000) ArticleTitleRegulation of source/sink relations by cytokinins Plant Growth Regul 32 359–367
F Rolland B Moore J Sheen (2002) Sugar sensing and signaling in plants Plant Cell S185–S205
JB Sherwood (1984) The chemistry and physiology of erythropoietin Vitamins and Hormones. Academic Press New York 161–211
GQ Tang M Lüscher A Sturm (1999) ArticleTitleAntisense repression of vacuolar and cell wall invertase in transgenic carrot alters early plant development and sucrose partitioning Plant Cell 11 177–189
T Werner V Motyka M Strnad T Schmüling (2001) ArticleTitleRegulation of plant growth by cytokinin Proc Natl Acad Sci USA 98 10487–10492
R Zhao V Dielen JM Kinet M Boutry (2000) ArticleTitleCosuppression of a plasma membrane H+-ATPase isoform impairs sucrose translocation, stomatal opening, plant growth, and male sterility Plant Cell 12 535–546
Author information
Authors and Affiliations
Corresponding author
Additional information
These authors contributed equally to this work
Rights and permissions
About this article
Cite this article
Cheon, B.Y., Kim, H.J., Oh, K.H. et al. Overexpression of human erythropoietin (EPO) affects plant morphologies: retarded vegetative growth in tobacco and male sterility in tobacco and Arabidopsis. Transgenic Res 13, 541–549 (2004). https://doi.org/10.1007/s11248-004-2737-3
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/s11248-004-2737-3