Inducible promoter containing JERE (Jasmonic Acid and Elicitor Responsive Element) box as well as construction method and application to genetic engineering

A promoter and inducible technology, applied in the field of plant genetic engineering, to achieve the effect of low expression, moderate expression intensity and long duration

Inactive Publication Date: 2010-11-17
FUJIAN AGRI & FORESTRY UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Constitutive promoters can be used to evaluate the biological function of a gene, the amount of transgene expression and its activity, but the main problems in the practical application of improving transgenic crops are: (1) the genes driven by constitutive promoters in various plant tissues It is expressed to varying degrees in the plant, which drives the expression of exogenous genes in various tissues and all developmental stages of the plant, and produces a large number of heterologous proteins or metabolites that accumulate in the plant, breaking the original metabolic balance of the plant and increasing the growth rate of the plant. Some products are even toxic to the growth and development of plants, causing a huge waste of material and energy, and at the same time changing the shape of plants, affecting the normal growth and development of plants, and even leading to death; (2) All cells of plants may All will enter the defense mode, even when there is no pathogen infection, the disease resistance response is still carried out; (3) the transgene product regulated by the constitutive promoter can accumulate in the plant tissues eaten by humans and animals, but the biosafety requirements cannot have transgenic product
However, direct evidence that elements such as the JERE box respond to pathogens or elicitors in rice and monocots is still lacking.

Method used

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  • Inducible promoter containing JERE (Jasmonic Acid and Elicitor Responsive Element) box as well as construction method and application to genetic engineering
  • Inducible promoter containing JERE (Jasmonic Acid and Elicitor Responsive Element) box as well as construction method and application to genetic engineering
  • Inducible promoter containing JERE (Jasmonic Acid and Elicitor Responsive Element) box as well as construction method and application to genetic engineering

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Embodiment Construction

[0034] 1.1.1.2 Biochemical reagents: DNA rapid recovery kit and plasmid extraction kit were purchased from Shanghai Sangon Bioengineering Co., Ltd., XbaI, SacI, SpeI, PfuTaq enzyme, rTaq Enzyme, T4 ligase and dNTP were purchased from TaKaRa Company, Gateway vector construction kit was purchased from Invitrogen Company, gentamicin, kanamycin, rifampicin, carbenicillin, hygromycin, X-gluc, etc. Product of Sigma Corporation. All other chemical reagents were domestic chemically pure.

[0035] 1.1.1.3 Primers: The primer sequences were synthesized by Shanghai Sangon Bioengineering Co., Ltd., as shown in Table 1.

[0036] Table 1 Primer name and nucleotide sequence

[0037]

[0038] 1.1.1.4 Sequence synthesis: According to the JERE box sequence in Ohme-TakagiandShinshi, (where the underlined bases are added, so that the double strands synthesized by annealing the positive and negative strands have wxya and SpeI Two restriction enzyme sites) were entrusted to Shanghai Boy...

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Abstract

The invention provides an inducible promoter containing a JERE (Jasmonic Acid and Elicitor Responsive Element) box as well as a construction method and the application to genetic engineering. The inducible promoter at least contains a nucleotide sequence shown as SEQ ID No.1. The construction method comprises the following step of: fusing a stress-response-stressed JERE box identified in dicotyledon parsley with a CaMV35S promoter core sequence to construct an inducible promoter. The inducible promoter containing the JERE box has low background expression, high response speed, long lasting time and suitable expression strength, can favorably response biological stress and abiotic stress, such as the invasion of rice blast fungus, the feeding of rice cnaphalocrocis medinalis, the mechanical damage, and the like in transgenic rice, has very important application values in the disease-resistant and pest-resistant genetic engineering of monocotyledon rice, and transgenic plant strains also have very important application values in the high throughput screening of small-molecule compounds for inducing plants to resist diseases.

Description

technical field [0001] The present invention relates to an inducible promoter and its construction and application in genetic engineering, and more specifically relates to the application of an inducible promoter containing a JERE box in the genetic engineering of monocotyledonous rice disease and pest resistance, belonging to the plant field of genetic engineering technology. Background technique [0002] The infection of pathogenic microorganisms such as fungi, bacteria, and viruses often causes a decline in the yield and quality of various crops. Breeding and promoting disease-resistant varieties is the safest and most effective way to prevent and control crop diseases and ensure stable and high yields of crops. Using the plant's own disease-resistant genes, many new disease-resistant varieties have been bred through conventional breeding and molecular marker-assisted breeding methods. However, the utilization of resistance genes among species has certain limitations....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/10A01H5/00
Inventor 何水林黄定全王小燕王芳权方开星刘志钦官德义牟少亮
Owner FUJIAN AGRI & FORESTRY UNIV
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