Abstract
A method is described to deliver mRNA to plant protoplasts using polyethylene glycol for transient expression. In order to optimize delivery and subsequent expression, we have examined a number of parameters that affect expression levels including the requirement for Mg2+ and Ca2+ ions, carrier nucleic acid, and the linearity of the dose-response. Delivery of mRNA using PEG results in levels of expression that are comparable to those obtained with electroporation. This method is particularly useful for those species in which electroporation does not work well. Moreover, the use of PEG to deliver mRNA is an inexpensive alternative to electroporation in that it does not require an electroporator.
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Abbreviations
- PEG:
-
Polyethylene glycol
- DTT:
-
dithiothreitol
- MES:
-
2(N-morpholino)ethanesulfonic acid
References
Bradford MM (1976) Anal Biochem 72: 248–254
Dawson JRO, Dickerson PE, King JM, Sakai F, Trim ARH, Watts JW (1978) Z Naturforsch C 33: 548–551
De Wet JR, Wood KV, DeLuca M, Helinski DR, Subramani S (1987) Mol Cell Biol 7: 725–737
Fromm M, Callis J, Taylor LP, Walbot V (1987) Meth Enzymol 153: 351–366
Gallie DR, Feder JN, Schimke RT, Walbot V (1991) Mol Gen Genet 228: 258–264
Gallie DR, Lucas WJ, Walbot V (1989) Plant Cell 1: 301–311
Gallie DR, Sleat DE, Watts JW, Turner PC, Wilson TMA (1987) Nucl Acids Res 15: 3257–3273
Krens FA, Molendijk L, Wullems GJ, Schilperoort RA (1982) Nature 296: 72–74
Maas C, Werr W (1989) Plant Cell Rep 8: 148–151
Melton DA, Krieg PA, Rebagliati MR, Maniatis T, Zinn K, Green MR (1984) Nucl Acids Res 12: 7035–7056
Negrutiu I, Shillito R, Potrykus I, Biasini G, Sala F (1987) Plant Mol Biol 8: 363–373
Samac DA, Nelson SE, Loesch-Fries LS (1983) Virol 131: 455–462
Tyagi S, Sporlein B, Tyagi AK, Herrmann RG, Koop HU (1989) Theor Appl Genet 78: 287–292
Vasil V, Hauptmann RM, Morrish FM, Vasil IK (1988) Plant Cell Rep 7: 499–503
Yisraeli JK, Melton DA (1989) Meth Enzymol 180: 42–50
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Communicated by F. Constabel
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Gallie, D.R. Introduction of mRNA to plant protoplasts using polyethylene glycol. Plant Cell Reports 13, 119–122 (1993). https://doi.org/10.1007/BF00235304
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DOI: https://doi.org/10.1007/BF00235304