Summary
Dicot and monocot chloroplast targeting peptides (CTPs) were evaluated for their effect on targeting, processing, and expression of two reporter proteins in maize cells. When tested transiently in maize leaf protoplasts, the maize ribulose bisphosphate carboxylase small subunit CTP required the inclusion of the amino terminus of mature small subunit protein to target β-glucuronidase (GUS) to the plastid. To remove this amino terminal extension from GUS after import and processing, a repeat of the native processing site was inserted between the native mature protein and the reporter protein. This repeat processing site was used with less efficiency than the native site. Parallel constructs using the Arabidopsis thaliana small subunit and maize granule-bound starch synthase CTPs also localized GUS, but varied in repeat site use and GUS expression levels. Data from the CTP fusions with GUS were generally confirmed with fusions to an allosteric variant of E. coli ADP-glucose pyrophosphorylase. Plastid targeting of this enzyme was required for starch enhancement of transgenic BMS cells.
Similar content being viewed by others
Abbreviations
- BMS:
-
maize Black Mexican Sweet suspension culture cells
- CTP:
-
chloroplast targeting peptide
- glgC16:
-
an allosteric variant of E. coli ADP-glucose pyrophosphorylase
- GUS:
-
β-glucuronidase
- LUX:
-
luciferase
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- SSU:
-
small subunit of ribulose bisphosphate carboxylase
References
Callis J, Fromm M, Walbot V (1987) Genes and Development 1: 1183–1200
Coe EH Jr, Neuffer MG, Hoisington DA (1988) In: GF Sprague, JW Dudley (eds) Corn and Corn Improvement, third ed. ASA, Inc, Madison, pp 81–258
Fromm ME, Morrish F, Armstrong C, Williams R, Thomas J, Klein TM (1990) Bio/Technology 8: 833–839
Jefferson RA (1987) Plant Mol Biol Reporter 5: 387–405
Kavanaugh TA, Jefferson RA, Bevan MW (1988) Mol Gen Genet 215: 38–45
Klein TM, Gradziel T, Fromm ME, Sanford JC (1989) Bio/Technology 6: 559–563
Klösgen RB, Saedler H, Weil J-H (1989) Mol Gen Genet 217: 155–161
Klösgen RB, Weil J-H (1991) Mol Gen Genet 225: 297–304
Ko K, Ko ZW (1992) J Biol Chem 267: 13910–13916
Kuntz M, Simons A, Schell J, Schreier PH (1986) Mol Gen Genet 205: 454–460
Leung P, Lee Y-M, Greenberg E, Esch K, Boylan S, Preiss J (1986) J Bacteriology 167: 82–88
Olins PO, Rangwala SH (1989) J Biol Chem 264: 16973–16976
Robinson SP (1987) Methods in Enzymol 148: 188–206
Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning. Cold Spring Harbor Press, NY
Sheen J (1990) Plant Cell 2: 1027–1038
Shure M, Wessler S, Federoff N (1983) Cell 35: 225–233
Stark DM, Timmerman KP, Barry GF, Preiss J, Kishore GM (1992) Science 258: 287–292
Wasmann CC, Reiss B, Bartlett SG, Bohnert HJ (1986) Mol Gen Genetics 205: 446–453
Author information
Authors and Affiliations
Additional information
Communicated by R. N. Beachy
Rights and permissions
About this article
Cite this article
Russell, D.A., DeBoer, D.L., Stark, D.M. et al. Plastid targeting of E. coli β-glucuronidase and ADP-glucose pyrophosphorylase in maize (Zea mays L.) cells. Plant Cell Reports 13, 24–27 (1993). https://doi.org/10.1007/BF00232309
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00232309