Abstract
The early events of transient gene expression have been investigated monitoring CAT activity in tobacco protoplasts encoded by the recombinant plasmid pRT101cat. The first appearance of CAT activity was observed within 30 minutes after the outset of cultivation, and maximal values were obtained between four and 24 hours. CAT expression, at the level of RNA synthesis, could not be inhibited by cordycepin (3′deoxyadenine) added one hour after protoplast plating, whereas cycloheximide, an inhibitor of protein synthesis, showed an influence during the first four hours. This indicates a rapid decay of biologically active forms of both the DNA transferred and the CAT-mRNA synthesized within the first hours. These results suggest that in the tobacco protoplast system CAT protein stability lasts up to two weeks rather than a continuous synthesis of new enzyme.
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Abbreviations
- BAP:
-
Benzylaminopurin
- CaMV:
-
Cauliflower Mosaic Virus
- CAT:
-
Chloramphenicolacetyltransferase
- PEG:
-
Polyethylenglycol
- NAA:
-
Naphtylaceticacid
Literature
Ballas N, Zakai N Loyter A (1987) Exp Cell Res 170: 228–234
Cline MG, Rehm MM, Wilson RH (1974) Plant Physiol 54: 160–163
Delseny M, Peralta MT, Guitton Y (1975) Biochem Biophys Res Com 64: 1278–1285
Ebert PR, Ha SB, An G (1987) Proc Natl Acad Sci 84: 5745–5749
Folger KR, Thomas K, Capecchi MR (1985) Mol Cell Biol 5: 59–69
French R, Janda M, Ahlquist P (1986) Science 231: 1294–1297
Fromm M, Taylor LP, Walbot V (1985) Proc Natl Acad Sci 82: 5824–5828
Gorman CM, Moffat LF, Howard BH (1982) Mol Cell Biol 2: 1044–1051
Hain R, Stabel P, Czernilofsky AP, Herrera-Estrella L, Schell J (1985) Mol Gen Genet 199: 161–168
Hauptmann RM, Ozias-Akins P, Vasil V, Tabaeizadeh Z, Rogers SG, Horsch RG, Vasil IK, Fraley RT (1987) Plant Cell Reports 6: 265–270
Junker B, Zimny J, Lührs R, Lörz H (1987) Plant Cell Reports 6: 329–332
Linsmaier EM, Skoog F (1965) Physiol Plant 18: 100–127
Loyter A, Scangos GA, Ruddle FH (1982) Proc Natl Acad Sci 79: 422–426
Maliga P, Sz.-Breznovitis A, Marton L (1973) Nature 244: 29–30
McMahon D (1975) Plant Physiol 55: 815–821
Nagy JI, Maliga P (1976) Z Pflanzenphysiol 78: 453–455
Okada K, Takebe I, Nagata T (1986) Mol Gen Genet 205:398–403
Palmer CE (1985) Plant Cell Physiol 26: 407–417
Pröls M, Schell J, Steinbiß H-H (1988) In: Electroporation and Electrofusion in Cell Biology (eds Neumann, Sowers, Jordan), in press
Töpfer R, Matzeit V, Gronenborn B, Schell J, Steinbiß H-H (1987) Nucl Acids Res 15: 5890
Velten J, Schell J (1985) Nucl Acids Res 13: 6981–6998
Werr W, Lörz H (1986) Mol Gen Genet 202: 471–475
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Pröls, M., Töpfer, R., Schell, J. et al. Transient gene expression in tobacco protoplasts: I. Time course of CAT appearance. Plant Cell Reports 7, 221–224 (1988). https://doi.org/10.1007/BF00272528
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DOI: https://doi.org/10.1007/BF00272528