Transient Expression of Chimaeric Genes in Dividing and Non-dividing Cereal Protoplasts After PEG-induced DNA Uptake

Overview

Journal Plant Cell Rep

Publisher Springer

Date 2013 Nov 20

PMID 24248837

Citations 20

Authors

B Junker

J Zimny

R Luhrs

H Lorz

Affiliations

Soon will be listed here.

Abstract

Transient expression of chimaeric genes (neomycin phosphotransferase fused to four different promoters) was detected in suspension culture derived protoplasts of maize, barley and rice, in mesophyll protoplasts of maize, rice, rye, and root protoplasts of maize. The introduction and expression of foreign genes could be performed with both dividing and non-dividing protoplasts by applying the PEG transformation method. The significance of this method for the functional analysis of genes was demonstrated by the differential expression of a regulated gene in protoplasts of different tissues in agreement with its expression in the donor tissue.

Citing Articles

Overexpression of GFP Fusions of Regulators of the SAC from Arabidopsis thaliana.

Pain C Methods Mol Biol. 2024; 2874:21-32.

PMID: 39614044 DOI: 10.1007/978-1-0716-4236-8_3.

Biolistic Approach for Transient Gene Expression Studies in Plants.

Lacroix B, Citovsky V Methods Mol Biol. 2020; 2124:125-139.

PMID: 32277451 PMC: 7217558. DOI: 10.1007/978-1-0716-0356-7_6.

Altered Promoter and G-Box Binding Factor for Gene Grown from Seeds after Spaceflight.

Gan L, Chao Y, Su H, Ren Y, Yin S, Han L Int J Mol Sci. 2019; 20(6).

PMID: 30901811 PMC: 6471272. DOI: 10.3390/ijms20061398.

Transient gene expression in tobacco protoplasts: II. Comparison of the reporter gene systems for CAT, NPT II, and GUS.

Topfer R, Prols M, Schell J, Steinbiss H Plant Cell Rep. 2013; 7(4):225-8.

PMID: 24241753 DOI: 10.1007/BF00272529.

Transient gene expression in tobacco protoplasts: I. Time course of CAT appearance.

Prols M, Topfer R, Schell J, Steinbiss H Plant Cell Rep. 2013; 7(4):221-4.

PMID: 24241752 DOI: 10.1007/BF00272528.

References

Velten J, Schell J . Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res. 1985; 13(19):6981-98. PMC: 322017. DOI: 10.1093/nar/13.19.6981. View

Springer B, Werr W, Starlinger P, Bennett D, Zokolica M, Freeling M . The Shrunken gene on chromosome 9 of Zea mays L is expressed in various plant tissues and encodes an anaerobic protein. Mol Gen Genet. 1986; 205(3):461-8. DOI: 10.1007/BF00338083. View

Graves A, Goldman S . The transformation of Zea mays seedlings with Agrobacterium tumefaciens. Plant Mol Biol. 2013; 7(1):43-50. DOI: 10.1007/BF00020130. View

Zimny J, Lorz H . Plant regeneration and initiation of cell suspensions from root-tip derived callus of Oryza sativa L. (rice). Plant Cell Rep. 2013; 5(2):89-92. DOI: 10.1007/BF00269241. View

Fromm M, Taylor L, Walbot V . Expression of genes transferred into monocot and dicot plant cells by electroporation. Proc Natl Acad Sci U S A. 1985; 82(17):5824-8. PMC: 390645. DOI: 10.1073/pnas.82.17.5824. View

Dudits D, Hadlaczky G, Levi E, Fejer O, Haydu Z, Lazar G . Somatic hybridisation of Daucus carota and D. capillifolius by protoplast fusion. Theor Appl Genet. 2013; 51(3):127-32. DOI: 10.1007/BF00273824. View

Ou-Lee T, Turgeon R, Wu R . Expression of a foreign gene linked to either a plant-virus or a Drosophila promoter, after electroporation of protoplasts of rice, wheat, and sorghum. Proc Natl Acad Sci U S A. 1986; 83(18):6815-9. PMC: 386600. DOI: 10.1073/pnas.83.18.6815. View

Pietrzak M, Shillito R, Hohn T, Potrykus I . Expression in plants of two bacterial antibiotic resistance genes after protoplast transformation with a new plant expression vector. Nucleic Acids Res. 1986; 14(14):5857-68. PMC: 311596. DOI: 10.1093/nar/14.14.5857. View

Reiss B, Sprengel R, Will H, Schaller H . A new sensitive method for qualitative and quantitative assay of neomycin phosphotransferase in crude cell extracts. Gene. 1984; 30(1-3):211-7. DOI: 10.1016/0378-1119(84)90122-7. View

10.

Potrykus I, Harms C, Lorz H . Callus formation from cell culture protoplasts of corn (Zea mays L.). Theor Appl Genet. 2013; 54(5):209-14. DOI: 10.1007/BF00267709. View

11.

Howard E, Walker J, Dennis E, Peacock W . Regulated expression of an alcohol dehydrogenase 1 chimeric gene introduced into maize protoplasts. Planta. 2013; 170(4):535-40. DOI: 10.1007/BF00402988. View

12.

Fromm M, Taylor L, Walbot V . Stable transformation of maize after gene transfer by electroporation. Nature. 1986; 319(6056):791-3. DOI: 10.1038/319791a0. View

13.

Schreier P, Seftor E, Schell J, Bohnert H . The use of nuclear-encoded sequences to direct the light-regulated synthesis and transport of a foreign protein into plant chloroplasts. EMBO J. 1985; 4(1):25-32. PMC: 554146. DOI: 10.1002/j.1460-2075.1985.tb02312.x. View