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WO-2026092652-A1 - CONSTITUTIVE PROMOTER AND USE THEREOF

WO2026092652A1WO 2026092652 A1WO2026092652 A1WO 2026092652A1WO-2026092652-A1

Abstract

Provided are a constitutive promoter and a use thereof. The nucleotide sequence of the constitutive promoter comprises SEQ ID NO: 34. The constitutive promoter shows activity in most tissues and many types of cells of plants, especially in roots, stems, leaves, filaments, pollen, seeds, and inflorescence apices of plants, and has a broad application prospect in plants.

Inventors

  • WANG, CHEN
  • ZHANG, MING
  • ZHAO, Pusheng
  • LIU, XIAOGUANG

Assignees

  • 北京大北农生物技术有限公司

Dates

Publication Date
20260507
Application Date
20251031
Priority Date
20241104

Claims (14)

  1. A constitutive promoter, characterized in that the nucleotide sequence of the constitutive promoter includes SEQ ID NO:34.
  2. The constitutive promoter according to claim 1, wherein the nucleotide sequence of the constitutive promoter includes SEQ ID NO:34, and the constitutive promoter is a truncated sequence of SEQ ID NO:1.
  3. The constitutive promoter according to claim 1 or 2, characterized in that the nucleotide sequence of the constitutive promoter is as shown in SEQ ID NO:1, SEQ ID NO:33, and SEQ ID NO:34.
  4. A chimeric gene, characterized in that it comprises a constitutive promoter according to any one of claims 1-3 operably linked to a target nucleic acid; Preferably, the target nucleic acid encodes the target protein.
  5. An expression cassette, characterized in that it comprises a constitutive promoter as described in any one of claims 1-3 or a chimeric gene as described in claim 4.
  6. A recombinant vector, characterized in that it comprises a constitutive promoter as described in any one of claims 1-3, a chimeric gene as described in claim 4, or an expression cassette as described in claim 6.
  7. A host cell, characterized in that it comprises a constitutive promoter as described in any one of claims 1-3, a chimeric gene as described in claim 4, an expression cassette as described in claim 5, or a recombinant vector as described in claim 6.
  8. A method for expressing a target nucleic acid in a plant, characterized by comprising: stably integrating the target nucleic acid, which is operatively linked to a constitutive promoter according to any one of claims 1-3, into a plant cell; Preferably, the target nucleic acid is constitutively expressed in plant tissues; Preferably, the target nucleic acid encodes the target protein; Preferably, the target nucleic acid encodes a herbicide-resistant protein or an insect-resistant protein.
  9. The method for expressing a target nucleic acid in a plant according to claim 8, wherein the plant is corn, Arabidopsis thaliana, rapeseed, tobacco, soybean, cotton, chili pepper, beet, pumpkin, eggplant, Chinese cabbage, carrot, tomato, pea, spinach, potato, or peanut.
  10. A plant or plant part, characterized in that it comprises the constitutive promoter of any one of claims 1-3, the chimeric gene of claim 4, the expression cassette of claim 5, or the recombinant vector of claim 6.
  11. A method for obtaining processed agricultural products, characterized by processing the harvest of the plant or part thereof as described in claim 10 to obtain processed agricultural products.
  12. The use of the constitutive promoter according to any one of claims 1-3 for constitutive expression of a target nucleic acid in plant tissues.
  13. The use of the constitutive promoter according to claim 12 for constitutively expressing a target nucleic acid in plant tissues, characterized in that the target nucleic acid encodes a target protein; Preferably, the target nucleic acid encodes a herbicide-resistant protein or an insect-resistant protein.
  14. The use of the constitutive promoter according to claim 12 or 13 for constitutively expressing the target nucleic acid in plant tissues, characterized in that the plant is maize, Arabidopsis thaliana, rapeseed, tobacco, soybean, cotton, pepper, beet, pumpkin, eggplant, Chinese cabbage, carrot, tomato, pea, spinach, potato or peanut.

Description

Constitutive promoters and their applications Technical Field This invention relates to a constitutive promoter and its use, and more particularly to a constitutive promoter derived from millet and its use. Background Technology One of the goals of plant genetic engineering is to modify plants according to human needs, thereby producing plants with desired characteristics or traits. Commonly desired traits include improved nutritional quality, increased yield, conferring resistance to pests and diseases, improved drought and stress tolerance, improved horticultural quality, and conferring herbicide resistance. Currently, researchers are able to obtain exogenous genes (such as heterologous or naturally derived genes) and integrate them into the plant genome for expression to exhibit the corresponding traits. Choosing appropriate regulatory signals is crucial for expressing target genes in plants. Typical regulatory signals include promoter regions, 5' untranslated guide sequences, and 3' transcription terminators/polyadenylated sequences. A promoter is a DNA sequence located upstream of the 5' end of a structural gene that can be recognized, bound, and initiate gene transcription by RNA polymerase. In plant genetic engineering, promoters are often classified into three categories based on their mode of action and function: constitutive promoters, specific promoters, and inducible promoters. Constitutive promoters are those that maintain continuous activity in most or all tissues and guide RNA synthesis at a relatively similar level. Based on their effectiveness in guiding RNA synthesis, they can be further divided into strong, moderate, and weak promoters. Constitutive promoters are particularly advantageous in situations where it is necessary to simultaneously express the target gene (or chimeric gene) in different plant tissues to obtain the desired gene function. Several constitutive promoters that function in plant cells have been described in existing literature. These promoters include the soybean cell elongation factor gene promoter Gm17gTsf1, the nos promoter carried on the Agrobacterium tumefaciens tumor-inducing plasmid, the octopus amino acid synthase (OCS) promoter, and cauliflower mosaic virus (CaMV) promoters, such as the CaMV 19S or 35S promoter, the CaMV 35S promoter with repeat enhancers, and the Scrophularia mosaic virus (FMV) 35S promoter. These promoters have been used in transgenic plant constructs. Although some constitutive promoters have been obtained, the discovery and isolation of more constitutive promoters that are safe, efficient, and capable of controlling the expression of target genes (or chimeric genes) at different levels or applied to the superimposed expression of multiple genes in the same transgenic plant remains a hot topic of ongoing interest in this field. Summary of the Invention The purpose of this invention is to provide a novel constitutive promoter and its uses. This promoter is derived from millet (Panicum miliaceum L.) and can regulate the efficient expression of target nucleic acids in plant tissues such as roots, stems, leaves, filaments, pollen, grains, and the tip of the rachis. It provides a new tool and option for plant genetic engineering and has a very broad application prospect. In a first aspect, the present invention provides a constitutive promoter having a nucleotide sequence comprising SEQ ID NO:34. Preferably, the constitutive promoter has a nucleotide sequence including SEQ ID NO:34, and the constitutive promoter is derived from SEQ ID NO:1. More preferably, the constitutive promoter has a nucleotide sequence including SEQ ID NO:34, and is a part or a fragment of SEQ ID NO:1. More preferably, the constitutive promoter has a nucleotide sequence including SEQ ID NO:34, and is a truncated sequence of SEQ ID NO:1. More preferably, the constitutive promoter has a nucleotide sequence including SEQ ID NO:34, and is a 5' (N-terminus) truncated sequence of SEQ ID NO:1. More preferably, the 5' end (N end) is truncated to no more than (less than or equal to) 1111 bp. More preferably, the constitutive promoter has the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:33, and SEQ ID NO:34. Secondly, the present invention also provides a chimeric gene comprising the above-described constitutive promoter operatively linked to a target nucleic acid. Preferably, the target nucleic acid encodes the target protein. Thirdly, the present invention also provides an expression cassette containing the constitutive promoter or chimeric gene of the present invention. Fourthly, the present invention also provides a recombinant vector comprising the constitutive promoter, chimeric gene, or expression cassette of the present invention. Fifthly, the present invention also provides a host cell comprising the constitutive promoter, chimeric gene, expression cassette, or recombinant vector of the present invention. In a sixth aspect, the present invention also provides a meth