Constitutive plant promoters

A promoter and plant technology, applied in plant products, botanical equipment and methods, angiosperms/flowering plants, etc., can solve problems such as activity changes and expression changes

Inactive Publication Date: 2009-02-18
KEYGENE NV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, a common disadvantage of existing constitutive promoters is that their activity often changes, such as organ or developmentally regulated expression or stress-induced expression changes

Method used

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  • Constitutive plant promoters
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0121] Example 1 - Materials and methods

[0122] Differential expression analysis of the common tomato variety (Lycopersicon esculentum) Moneyberg using cDNA.

[0123] The common tomato variety (Lycopersicon esculentum) Moneyberg was tested for expression under stress conditions.

[0124] Transformation in tobacco (Nicotiana tabacum) variety SR1 and common tomato variety RZ 52201 was performed using vectors pKG8136, pKG8137 and pKG1700. Plant transformations were performed as described in the following references:

[0125] Tomato and Tobacco Transformation:

[0126] An G., B.D. Watson and C.C. Chiang. 1986. Transformation of tobacco, tomato, potato, and Arabidopsis thaliana using a binary Ti vector system. Plant Physiol. 81: 301-305.

[0127] Tobacco transformation: Horsch R.B., J. Fry, N. Hoffman, J. Neidermeyer, S.G. Rogers and R.T. Fraley. 1988. Leaf disc transformation. Plant Molecular Biology Manual A5. Dordrecht, Netherlands, Kluwer Academic Publishers. pp. 1-9.

...

Embodiment 2

[0139] Example 2 Selection of genes for isolating promoters

[0140] use (Volkmuth W., et al., 2003, Genome-Wide Analysis of the Arabidopsis Transcriptome, OMICS, 7, 2; Vos, P. and Stanssens P., 2002, AFLP-based transcript profiling, Current Protocols in Molecular Biology, unit 25B.5., Ausubel, F.M., Brent, R., Kingston, R.E., Moore, D.D., Seidman, J.G. Smith, J.A., Struhl, K., John Wiley and Sons, New York; and Vos et al., 1995, AFLP: a new technique for DNA fingerprinting, Nucleic Acids Research 23:4407-4414) Tomato plant material from various developmental stages (young leaves, old leaves, stems, roots, detached leaves, detached stems, detached roots, calluses, immature and mature green fruits, reddened fruits, red fruits, Differential analysis of the cDNA of flowers and total seedlings). Expression profiles were generated using all possible Taq1 / Mse1 primers in combination with two selective nucleotides of the Taq1 primer and three selective nucleotides of the Mse1 ...

Embodiment 3

[0151] Example 3 Isolation of AA6 promoter sequence

[0152] Linker PCR (linker PCR) was performed using genomic DNA from BAC9 as a template with primers designed for a derivative sequence of a tomato gene named cysteine ​​synthase (see below for primer sequence). The 5' and 3' sequences of the cysteine ​​synthase UTR were obtained. Using the genomic DNA of Moneyberg tomato as a template, the 3'UTR sequence (SEQ ID NO: 3) was obtained by long-range PCR (long range PCR).

[0153] Primers designed against the cysteine ​​synthase sequence:

[0154] For promoter 5′-GTTCGATGAGGACACTCTCGC-3

[0155] and nested primer 5′-CAATTAAAGTTGCTAAGCGTCCTGA-3′

[0156] To terminator 5′-TCAGTTACATCCTTGGCAATTCC-3′

[0157] and nested primer 5′-CAGAGAACATGACTGTGGAGCC-3′

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Abstract

Strong, constitutive plant promoters are provided, referred herein to as AA6 promoters, which remain strong and constitutive under biotic and / or abiotic stress conditions. Also provided are transgenic cells and organisms, especially plant cell and plants, comprising an AA6 promoter and methods for expressing nucleic acid sequences in cells and organisms using AA6 promoters.

Description

technical field [0001] The present invention relates to a class of strong constitutive plant promoters and their use. The promoters can be used for the expression of homologous or heterologous proteins in plants or plant cells, or for the expression of active nucleic acid molecules such as sense RNA and / or antisense RNA. The present invention provides nucleic acid sequences having promoter activity as well as chimeric genes, vectors and recombinant (transgenic) cells and organisms comprising them. The invention also provides methods for producing transgenic cells and organisms, especially plants and plant cells, comprising said promoters. Furthermore, the present invention provides cytoplasmic cysteine ​​synthase proteins and nucleic acid sequences encoding them. Background technique [0002] A wide variety of plant promoters are known in the art and are useful tools for expressing proteins or peptides in transgenic plants or plant cells or for silencing a gene or gene fam...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/82C12N9/88A01H5/00
CPCC12N15/8216C12N9/88C12N9/1088C12Y205/01047
Inventor M·T·J·德鲍思N·E·M·克瓦埃德福里格B·G·J·费尔兰斯-昂斯腾克L·K·瓦乔斯基
Owner KEYGENE NV
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