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Phytophthora induced artificially synthesized promoter PMP2 as well as recombinant expression vector and application thereof

An expression vector and promoter technology, applied in the application of recombinant expression vectors, Phytophthora inducible synthetic promoter PMP2 and its recombinant expression vector, Phytophthora inducible promoter field, can solve Phytophthora disease control methods complex, The control effect is not obvious, the lack of pathogen-inducible promoters and other problems

Active Publication Date: 2016-08-17
江苏南京农大科技开发有限责任公司
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  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to solve the problems in the prior art that the control method for Phytophthora disease is complicated, the control effect is not obvious, and the novel disease-resistant genetic engineering technology lacks available pathogen-inducible promoters, and provides a Phytophthora inducible promoter. Promoter

Method used

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  • Phytophthora induced artificially synthesized promoter PMP2 as well as recombinant expression vector and application thereof
  • Phytophthora induced artificially synthesized promoter PMP2 as well as recombinant expression vector and application thereof
  • Phytophthora induced artificially synthesized promoter PMP2 as well as recombinant expression vector and application thereof

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Embodiment 1

[0033] The acquisition of embodiment 1 novel cis element ATCCAA

[0034] 1. Screening of Phytophthora-specific inducible gene promoter structural elements based on microarray data

[0035] According to the microarray data of three different soybean varieties: V71-370, VPRIL9, and Sloan infected by Phytophthora sojae (Zhou L, Mideros SX, Bao L, et al. Infection and genotype remodel the entire soybean transcriptome[J].BMC Genomics, 2009,10(1):49-59.), analyzed the promoter regions of 180 genes whose expression was upregulated after Phytophthora soybean infection. We first collected some reported action element sequences, looked up the frequency and position distribution of these sequences in the 180 genes, and compared the expression of promoters containing these elements 1 day after inoculation with those without these elements. promoters for comparison. The positional distribution of these elements within the respective promoters was then analyzed. For the selection of new st...

Embodiment 2

[0039] Example 2 The artificially synthesized PMP2 promoter is a Phytophthora inducible promoter

[0040] 1. Synthesis of artificial promoter and vector construction

[0041] The new cis-element ATCCAA obtained by bioinformatics analysis, and its upstream and downstream flanking sequences of 12bp, a total of 30bp, artificially synthesized four repeating units of these 30bp, as the sequence of the artificially synthesized promoter PMP2 (such as SEQID NO.5 shown), used to construct plant expression vectors.

[0042] According to Chai C, Lin Y, Shen D, et al. Identification and Functional Characterization of the Soybean GmaPPO12 Promoter Conferring Phytophthora sojae Induced Expression [J]. PloS One, 2013, 8(6): 1-6. Construction of promoter fusion in the article The plant expression vector method of the GUS reporter gene obtains the PMP2::GUS recombinant plasmid fused with the GUS reporter gene by four repeating units, and the construction pattern of the vector is as follows ...

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Abstract

The invention belongs to the biotechnology field and discloses a phytophthora induced artificially synthesized promoter PMP2 as well as a recombinant expression vector and application thereof. The promoter takes an ATCCAA cis element and flanking sequences with the size of 12bp respectively at the upstream side and downstream side of the cis element as a core sequence. The cis element and flanking sequences, with the size of 12bp respectively, of the cis element in the upstream side and downstream side of a gene promoter are taken, namely that a sequence with the size of 30bp in total is taken, a tetramer is artificially synthesized as a promoter PMP2, the PMP2 is fused with a GUS report gene to be constructed into a plant expression vector pMDC162, and by virtue of an agrobacterium-medicated tobacco transient expression system, the pathogenic bacterium induced expression characteristic of the promoter is analyzed. Results show that the artificially synthesized PMP2 promoter can rapidly and efficiently drive the up-regulated induced expression of the GUS report gene under the induction of phytophthora. Therefore, the artificially synthesized PMP2 promoter is a pathogen induced promoter and provides a valuable pathogen induced promoter for plant anti-phytophthora genetic engineering.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Phytophthora inducible artificially synthesized promoter PMP2 and its recombinant expression vector, and the application of the Phytophthora inducible promoter and the recombinant expression vector. Background technique [0002] Effective control of plant diseases has always been a core issue in phytopathology research. For a long time, the use of chemical pesticides has been the main technology for disease control, but the problems faced in practical application are: on the one hand, there is a lack of effective pesticides, on the other hand, there are problems such as phytotoxicity and environmental pollution during the process of using pesticides. Later, a disease-resistant breeding strategy was proposed, but it was difficult to popularize and apply due to a series of problems such as long cycle, high cost, large scale and low precision. With the development of molecular biology ...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82A01H5/00
CPCC12N15/113C12N15/8222C12N15/8282C12N2310/10
Inventor 窦道龙柴春月曾文韬
Owner 江苏南京农大科技开发有限责任公司
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