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A kind of artificial chimeric promoter and its construction method

A technology of chimeric promoters and construction methods, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, genetic engineering, etc., and can solve the theoretical basis of DNA-protein interaction network and signal transduction pathways. Clear and other issues, to achieve the effect of low background activity, high expression intensity, and wide induction factors

Inactive Publication Date: 2021-12-31
CHENGDU UNIV
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Problems solved by technology

[0004] (1) At present, there is no technical means to control the precise regulation of disease resistance genes in the state of plant root pathogen infection
[0005] (3) There is no technical means to solve the effective connection mode of plant root-specific inducible promoter elements and wound-inducible specific promoter elements
[0006] The difficulty and significance of solving the above technical problems: Since the theoretical basis of the DNA-protein interaction network and signal transduction pathways is not very clear, the construction of artificial promoters is still facing severe challenges

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  • A kind of artificial chimeric promoter and its construction method
  • A kind of artificial chimeric promoter and its construction method
  • A kind of artificial chimeric promoter and its construction method

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Embodiment Construction

[0053]In order to make the object, technical solution and advantages of the present invention more clear, the present invention will be further described in detail below in conjunction with the examples. It should be understood that the specific embodiments described here are only used to explain the present invention, not to limit the present invention.

[0054] The present invention constructs a new artificial promoter by combining different promoter elements, replacing or redesigning.

[0055] Such as figure 1 As shown, the construction method of the artificial chimeric promoter provided by the embodiment of the present invention comprises the following steps:

[0056] S101: Splicing pTobR and pWIN into a TRWI (pTobR-pWin) promoter by fusion PCR, and inserting an artificial combination element C1 into the spliced ​​chimeric TRWI promoter to construct another chimeric promoter TC1W (pTobR -C1-pWin);

[0057] S102: Construct the chimeric promoter TC1W into the pBI121 binar...

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Abstract

The invention belongs to the technical field of genetic engineering, and discloses an artificial chimeric promoter and its construction method; pTobR and pWIN are combined by fusion PCR and an artificial combination element C1 is inserted therein to construct a chimeric promoter TC1W; The chimeric promoter TC1W was constructed into the pBI121 binary plant expression vector, and TC1W was transferred into tobacco through Agrobacterium-mediated transformation; GUS tissue staining and GUS protease activity assay were performed on the transgenic tobacco plants to explore the expression pattern of the downstream GUS gene driven by the fusion promoter TC1W . The present invention constructs a new artificial promoter by combining different promoter elements, replacing or redesigning it, and constructs a new promoter with the characteristics of wide induction factors, low background activity, high expression intensity, and fast expression initiation. Better control over the expression of downstream genes of interest. In the absence of inducing factors, the TC1W promoter only showed root specificity.

Description

technical field [0001] The invention belongs to the technical field of genetic engineering, and in particular relates to an artificial chimeric promoter and a construction method thereof. Background technique [0002] At present, the existing technologies commonly used in the industry are as follows: At present, the two most commonly used methods for constructing artificial chimeric promoters are the combination of core promoters and cis-elements and the bidirectional combination of unidirectional promoters. In addition, cis-acting elements from different sources are directly connected in series. Generally, when genetically improving crops, it is often hoped that the inserted exogenous gene can be expressed in a specific tissue at a specific developmental stage or under specific environmental conditions, so as to reduce the impact of strong activation of the gene on the normal growth of the crop. Although a large number of cis-acting elements and their corresponding transcr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82A01H5/00A01H6/82
CPCC12N15/8216
Inventor 杨星勇方小艳谢成建
Owner CHENGDU UNIV
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