Summary
Chimeric gene constructs were made in which two reporter genes, the neo and bar genes, encoding neomycin phosphotransferase II and phosphinothricin acetyl transferase, respectively, were placed under the control of the promoter of ats1A, one of four genes encoding the ribulose-1,5-bisphosphate carboxylase (Rubisco) small subunit (SSU) in Arabidopsis thaliana. In one set of constructs the fusions were made at the initiation codons, while in the second set the sequences encoding the ats1 A transit peptide were included. Significantly higher steady-state levels of RNA and protein were observed in leaves of transgenic plants varrying the latter constructions. Individual transgenic plants varied in their degree of tissue specific expression of the chimeric genes as well as in absolute levels of expression. Preliminary results suggest that the ats1 A promoter may be only weakly responsive to phytochrome.
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References
An G (1986) Development of plant promoter expression vectors and their use for analysis of differential activity of the nopaline synthase promoter in transformed tobacco cells. Plant Physiol 81:86–91
Beck E, Ludwig G, Auerswald EA, Reiss B, Schaller H (1982) Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene 19:327–336
Bennett J, Jenkins GI, Hartley MR (1984) Differential regulation of the accumulation of the light-harvesting chlorophyll a/b complex and ribulose bisphosphate carboxylase/oxygenase in greening pea leaves. J Cell Biochem 25:1–13
Cashmore AR (1982) The isolation of polyA+ messenger RNA from higher plants. In: Edelman M, Hallick RB, Chua N-H (eds) Methods in chloroplast molecular biology. Elsevier Biomedical Press, Amsterdam, pp 387–392
Cornelissen M, Vandewiele M (1989) Nuclear transcriptional activity of the tobacco plastid psbA promoter. Nucleic Acids Res 17:19–29
Coruzzi G, Broglie R, Edwards C, Chua N-H (1984) Tissue-specific and light-regulated expression of pea nuclear gene encoding the small subunit of ribulose-1,5-bisphosphate carboxylase. EMBO J 3:1671–1679
Dean C, van den Elzen P, Tamaki S, Dunsmuir P, Bedbrook J (1985) Differential expression of the eight genes of the petunia ribulose bisphosphate carboxylase small subunit multi-gene family. EMBO J 4:3055–3061
Deblaere R, Bytebier B, De Greve H, Deboeck F, Schell J, Van Montagu M, Leemans J (1985) Efficient octopine Ti plasmid-derived vectors for Agrobacterium-mediated gene transfer to plants. Nucleic Acids Res 13:4777–4788
Deblaere R, Reynaerts A, Hofte H, Hernalsteen J-P, Leemans J, Van Montagu M (1987) Vectors for cloning in plant cells. Methods Enzymol 153:277–292
De Block M, Botterman J, Vandewiele M, Dockx J, Thoen C, Gossele V, Movva NR, Thompson C, Van Montagu M, Leemans J (1987) Engineering herbicide resistance in plants by expression of a detoxifying enzyme. EMBO J 6:2513–2518
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21
Ellis RJ (1981) Chloroplast proteins: Synthesis, transport, and assembly. Annu Rev Plant Physiol 32:111–137
Fluhr R, Chua N-H (1986) Developmental regulation of two genes encoding ribulose-bisphosphate carboxylase small subunit in pea and transgenic Petunia plants: Phytochrome response and blue-light induction. Proc Natl Acad Sci USA 83:2358–2362
Fluhr R, Moses P, Morelli G, Coruzzi G, Chua N-H (1986) Expression dynamics of the pea rbcS multigene family and organ distribution of transcripts. EMBO J 5:2063–2071
Fraley RT, Rogers SG, Horsch RB (1986) Genetic transformation in higher plants. CRC Crit Rev Plant Sci 4:1–46
Gidoni D, Bond-Nutter D, Brosio P, Jones J, Bedbrook J, Dunsmuir P (1988) Coordinated expression between two photosynthetic petunia genes in transgenic plants. Mol Gen Genet 211:507–514
Harpster M, Townsend JA, Jones JDG, Bedbrook J, Dunsmuir P (1988) Relative strengths of the 35S cauliflower mosaic virus, 1′, 2′, and nopaline synthase promoters in transformed tobacco sugarbeet and oilseed rape callus tissue. Mol Gen Genet 212:182–190
Herrera-Estrella L, Depicker A, Van Montagu M, Schell J (1983) Expression of chimaeric genes transferred into plant cells using a Ti-plasmid-derived vector. Nature 303:209–213
Herrera-Estrella L, Van den Broeck G, Maenhaut R, Van Montagu M, Schell J, Timko M, Schell J, Timko M, Cashmore A (1984) Light-inducible and chloroplast-associated expression of a chimaeric gene introduced into Nicotiana tabacum using a Ti plasmid vector. Nature 310:115–120
Jones JDG, Dunsmuir P, Bedbrook J (1985) High level expression of introduced chimaeric genes in regenerated transformed plants. EMBO J 4:2411–2418
Jones JDG, Dean C, Gidoni D, Gilbert D, Bond-Nutter D, Lee R, Bedbrook J, Dunsmuir P (1988) Expression of bacterial chitinase protein in tobacco leaves using two photosynthetic gene promoters. Mol Gen Genet 212:536–542
Kaufman LS, Thompson WF, Briggs WR (1984) Differential red light requirements for phytochrome induced accumulation of cab RNA and rbcS RNA. Science 226:1447–1449
Kay R, Chan A, Daly M, McPherson J (1987) Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236:1299–1302
Koziel MG, Adams TL, Hazlet MA, Damm D, Miller J, Dahlbeck D, Jayne S, Staskawicz BJ (1984) A cauliflower mosaic virus promoter directs expression of kanamycin resistance in morphogenic transformed plant cells. J Mol Appl Genet 2:549–562
Krebbers E, Seurinck J, Herdies L, Cashmore AR, Timko MP (1988) Four genes in two diverged subfamilies encode the ribulose-1,5-bisphosphate carboxylase small subunit polypeptides of Arabidopsis thaliana. Plant Mol Biol 11:745–759
Kuhlemeier C, Green PJ, Chua N-H (1987) Regulation of gene expression in higher plants. Annu Rev Plant Physiol 38:221–257
Kung SD (1976) Tobacco fraction I protein: A unique genetic marker. Science 191:429–434
Lutcke HA, Chow KC, Mickel FS, Moss KA, Kern HF, Scheele GA (1987) Selection of AUG initation codons differs in plants and animals. EMBO J 6:43–48
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: a laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, New York
Manzara T, Gruissem W (1988) Oganization and expression of the gene encoding ribulose-1,5-bisphosphate carboxylase in higher plants. Photosynth Res 16:117–139
Maxam A, Gilbert W (1980) Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol 65:499–560
Meinkoth J, Wahl G (1984) Hybridization of nucleic acids immobilized on solid supports. Anal Biochem 138:267–284
Morelli G, Nagy F, Fraley RT, Rogers SG, Chua N-H (1985) A short conserved sequence is involved in the light-inducibility of a gene encoding ribulose 1,5-bisphosphate carboxylase small subunit of pea. Nature 315:200–204
Nagy F, Morelli G, Fraley RT, Rogers SG, Chua N-H (1985) Photoregulated expression of a pea rbcS gene in leaves of transgenic plants. EMBO J 4:3063–3068
Odell JT, Nagy F, Chua N-H (1985) Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313:810–812
Pruitt RE, Meyerowitz EM (1986) Characterization of the genome of Arabidopsis thaliana. J Mol Biol 187:169–183
Reiss B, Sprengel R, Schaller H (1984a) Protein fusions with the kanamycin resistance gene from transposon Tn5. EMBO J 3:3317–3322
Reiss B, Sprengel R, Will H, Schaller H (1984b) A new sensitive method for quantitative and qualitative assay of neomycin phosphotransferase in crude cell extract. Gene 30:211–218
Rethy R, Dedonder A, Fredericq H, De Greef J (1983) Factors affecting the induction and release of secondary dormancy in Kalanchoe seeds. Plant Cell Environ 6:731–738
Sanders PR, Winter JA, Barnason AR, Rogers SG, Fraley RT (1987) Comparison of cauliflower mosaic virus 35S and nopaline synthase promoters in transgenic plants. Nucleic Acids Res 15:1543–1558
Sazaki Y, Sakihama T, Kamikubo T, Shinozaki K (1983) Phytochrome-mediated regulation of two mRNAs, encoded by nuclei and chloroplasts of ribulose-1,5-bisphosphate carboxylase/oxygenase. Eur J Biochem 133:617–620
Shah DM, Tumer NE, Fischhoff DA, Horsch RB, Rogers SG, Fraley RT, Jaworski EG (1987) The introduction and expression of foreign genes in plants. Biotechnol Genet Eng Rev 5:81–106
Simpson J, Van Montagu M, Herrera-Estrella L (1986) Photosynthesis-associated gene families: differences in response to tissue-specific and environmental factors. Science 233:34–38
Sugita M, Gruissem W (1987) Developmental, organ-specific, and light-dependent expression of the tomato ribulose-1,5-bisphosphate carboxylase small subunit gene family. Proc Natl Acad Sci USA 84:7104–7108
Thompson CJ, Movva NR, Tizard R, Crameri R, Davies JE, Lawereys M, Botterman J (1987) Characterization of the herbicideresistance gene bar from Streptomyces hygroscopicus. EMBO J 6:2519–2523
Thompson WF, Everett M, Polans NO, Jorgensen RA, Palmer JD (1983) Phytochrome control of RNA levels in developing pea and mung-bean leaves. Planta 158:487–500
Timko MP, Herdies L, de Almeida E, Cashmore AR, Leemans J, Krebbers E (1988) Genetic engineering of nuclear-encoded components of the photosynthetic apparatus in Arabidopsis. In: Phillips M, Shoemaker S, Ottenbrite RM, Middlekauff RD (eds) The impact of chemistry on biotechnology. American Chemical Society Books Inc, Washington DC, pp 279–295
Tobin EM (1981) Phytochrome-mediated regulation of mRNAs for the small subunit of ribulose-1,5-bisphosphate carboxylase and the light-harvesting chlorophyll a/b-protein in Lemna gibba. Plant Mol Biol 1:34–51
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheet: Procedure and some applications. Proc Natl Acad Sci USA 76:4350–4354
Tumer NE, O'Connell KM, Nelson RS, Sanders PR, Beachy RN, Fraley RT, Shah DM (1987) Expression of alfalfa mosaic virus coat protein gene confers cross-protection in transgenic tobacco and tomato plants. EMBO J 6:1181–1188
Van den Broeck G, Timko MP, Kausch AP, Cashmore AR, Van Montagu M, Herrera-Estrella L (1985) Targeting of a foreign protein to chloroplasts by fusion to the transit peptide from the small subunit of ribulose-1,5-bisphosphate carboxylase. Nature 313:358–363
Velten J, Schell J (1985) Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res 13:6981–6999
Velten J, Velten L, Hain R, Schell J (1984) Isolation of a dual promoter fragment from the Ti plasmid of Agrobacterium tumefaciens. EMBO J 12:2723–2730
Willmitzer L (1988) The use of transgenic plants to study plant gene expression. Trends Genet 4:13–18
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De Almeida, E.R.P., Gossele, V., Muller, C.G. et al. Transgenic expression of two marker genes under the control of an Arabidopsis rbcS promoter: Sequences encoding the Rubisco transit peptide increase expression levels. Mol Gen Genet 218, 78–86 (1989). https://doi.org/10.1007/BF00330568
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DOI: https://doi.org/10.1007/BF00330568