Summary
In this paper we report on the use of a bidirectional enhancer cloning vehicle to isolate and characterize new enhancer sequences from Arabidopsis thaliana. A library of A. thaliana genomic Sau3A segments was constructed in Escherichia coli in the binary plasmid enhancer cloning vehicle pROA97. The T-DNA based vector carries abbreviated TATA regions from the cauliflower mosaic virus 35S transcription unit upstream of two genes. The library was transferred via triparental mating into Agrobacterium tumefaciens. The neomycin phosphotransferase II gene was used for selection of kanamycin-resistant transformed tobacco callus cells. Approximately 1100 transgenic plants were regenerated and assayed for expression of the E. coli β-glucuronidase (GUS) gene in leaves, stems, roots, or seeds. Plasmids carrying putative enhancer sequences were rescued from the genomes of transgenic plants and the cloned sequences were assayed for enhancer function in genetic selection experiments. Plants were regenerated from the kanamycin-resistant calli obtained in the secondary transformation experiments. Histochemical analysis of GUS activity in the leaf, stem, and root tissues of transgenic plants showed a variety of expression patterns. The DNA sequences are presented of five Arabidopsis segments which confer enhancer function.
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References
An G, Friesen JD (1979) Plasmid vehicles for direct cloning of Escherichia coli promoters. J Bacteriol 140:400–407
Andre D, Colau D, Schell J, Van Montagu M, Hernalsteens JP (1986) Gene tagging in plants by a T-DNA insertion mutagen that generates APH(3′)II-plant gene fusions. Mol Gen Genet 204:512–518
Bellen HJ, O'Kane CJ, Wilson C, Grossniklaus U, Pearson RK, Gehring WJ (1989) P-element-mediated enhancer detection: a versatile method to study development in Drosophila. Genes Dev 3:1288–1300
Benfey PN, Ren L, Chua N-H (1989) The CaMV 35S enhancer contains at least two domains which can confer different developmental and tissue-specific expression patterns. EMBO J 8:2195–2202
Bier E, Vaessin H, Shepard S, Lee K, McCall K, Barbel S, Ackerman L, Carretto R, Uemura T, Grell E, Jan LY, Jan YN (1989) Searching for pattern and mutation in the Drosophila genome with a P-lacZ vector. Genes Dev 3:1273–1287
Casadaban MJ, Cohen SN (1980) Analysis of gene control signals by DNA fusion and cloning in Escherichia coli. J Mol Biol 138:179–207
Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA minipreparation: version II. Plant Mol Biol Rep 1:19–21
Depicker A, Stachel S, Dhaese P, Zambryski P, Goodman HM (1982) Nopaline synthase: transcript mapping and DNA sequence. J Mol Appl Genet 1:561–573
Depicker A, Herman L, Jacobs A, Schell J, Van Montagu M (1985) Frequencies of simultaneous transformation with different TDNAs and their relevance to the Agrobacterium/ plant cell interaction. Mol Gen Genet 201:477–484
Ditta G, Stanfield S, Corbin D, Helinski DR (1980) Broad host range DNA cloning system for Gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci USA 77:7347–7351
Donnelly CE, Sonenshein AL (1982) Genetic fusion of the E. coli lacZ gene to a B. subtilis promoter. In: Ganesan AT, Chang S, Hoch JA (eds) Molecular cloning and gene regulation in Bacilli. Academic Press, New York, pp 63–72
Donnelly CE, Sonenshein AL (1984) Promoter-probe plasmid for Bacillus subtilis. J Bacteriol 157:965–967
Feitelson JS (1988) An improved plasmid for the isolation and analysis of Streptomyces promoters. Gene 66:159–162
Fluhr R, Moses P, Morelli G, Caruzzi G, Chua N-H (1986) Expression dynamics of the pea rbcS multigene family and organ distribution of the transcripts. EMBO J 5:2063–2071
Fraley RT, Rogers SG, Horsch RB, Sanders PR, Flick JS, Adams SP, Bittner ML, Brand LA, Fink CL, Fry JS, Gallupi GR, Goldberg SB, Hoffman NL, Woo SC (1983) Expression of bacterial genes in plant cells. Proc Natl Acad Sci USA 80:4803–4807
Gidoni D, Bond-Nutter D, Brosio P, Jones J, Bedbrook J, and Dunsmuir P (1988) Coordinated expression between two photosynthetic petunia genes in transgenic plants. Mol Gen Genet 211:507–514
Gossler A, Joyner AL, Rossant J, Skarnes WC (1989) Mouse embryonic stem cells and reporter constructs to detect developmentally regulated genes. Science 244:463–465
Herman LMF, Van Montagu MC, Depicker AG (1986) Isolation of tobacco DNA segments with plant promoter activity. Mol Cell Biol 6:4486–4492
Horinouchi S, Beppu T (1985) Construction and application of a promoter-probe plasmid that allows chromogenic identification in Streptomyces lividans. J Bacteriol 162:406–412
Horsch RB, Fry JE, Hoffman NL, Eichholtz D, Rogers SG, Fraley RT (1985) A simple and general method for transferring genes into plants. Science 227:1229–1231
Jefferson RA (1987) Assaying chimeric genes in plants: The GUS gene fusion system. Plant Mol Biol Rep 5:387–405
Jefferson RA, Kavanagh TA, Bevan MW (1987) GUS fusions: β-glucuronidase as a sensative and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
Klee HJ, Hayford MB, Rogers SG (1987) Gene rescue in plants: A model system for “shotgun” cloning by retransformation. Mol Gen Genet 210:282–287
Koncz C, Martini N, Mayerhofer R, Koncz-Kalman Z, Korber H, Redei GP, Schell J (1989) High frequency T-DNA-mediated gene tagging in plants. Proc Natl Acad Sci USA 86:8467–8471
Leutwiler LS, Hough-Evans BR, Meyerowitz EM (1984) The DNA of Arabidopsis thaliana. Mol Gen Genet 194:15–23
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning: A laboratory manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Maniatis T, Goodourn S, Fischer JA (1987) Regulation of inducible and tissue-specific gene expression. Science 236:1237–1245
O'Kane CJ, Gehring WJ (1987) Detection in situ of genomic regulatory elements in Drosophila. Proc Natl Acad Sci USA 84:9123–9127
O'Kane CJ, Stephens MA, McConnell D (1986) Integrable α-amylase plasmid for generating random transcriptional fusions in Bacillus subtilis. J Bacteriol 168:973–981
Ott RW, Ren L, Chua N-H (1990) A bidirectional enhancer cloning vehicle for higher plants. Mot Gen Genet 221:121–124
Petit A, Tempe J, Kerr A, Holsters M, Van Montagu M, Schell J (1978) Substrate induction of conjugative activity of Agrobacterium tumefaciens Ti plasmids. Nature 271:570–572
Pribnow D (1979) Genetic control signals in DNA. In: Goldberger RE (ed) Biological regulation and development. Plenum, New York, pp 219–277
Scherer S, Mann C, Davis RW (1982) Reversion of a promoter deletion in yeast. Nature 298:815–819
Simoens C, Alliotte TH, Mendel R, Muller A, Schiemann J, Van Lijsebettens M, Schell J, Van Montagu M, Inze D (1986) A binary vector for transferring genomic libraries to plants. Nucleic Acids Res 14:8073–8090
Teeri TH, Herrera-Estrella L, Depicker A, Van Montagu M, Palva ET (1986) Identification of plant promoters in situ by T-DNA-mediated transcriptional fusions to the nptII gene. EMBO J 5:1755–1760
Velten J, Schell J (1985) Selection-expression plasmid vectors for use in genetic transformation of higher plants. Nucleic Acids Res 13:6981–6998
Zambryski P (1989) Agrobacterium-plant cell DNA transfer. In: Berg DE, Howe MM (eds) Mobile DNA. American Society for Microbiology Washington DC, pp 309–333
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Communicated by H. Saedler
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Ott, R.W., Chua, NH. Enhancer sequences from Arabidopsis thaliana obtained by library transformation of Nicotiana tabacum . Molec. Gen. Genet. 223, 169–179 (1990). https://doi.org/10.1007/BF00265050
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DOI: https://doi.org/10.1007/BF00265050