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A Phytophthora inducible synthetic promoter pmp1 and its recombinant expression vector and application

An expression vector and induced expression technology, applied in the application of recombinant expression vectors, Phytophthora-inducible synthetic promoter PMP1 and its recombinant expression vector, Phytophthora-inducible promoter field, can solve the lack of pathogen-inducible promoters, Phytophthora disease control methods are complicated and the control effect is not obvious

Active Publication Date: 2019-04-19
江苏南京农大科技开发有限责任公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • 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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  • A Phytophthora inducible synthetic promoter pmp1 and its recombinant expression vector and application
  • A Phytophthora inducible synthetic promoter pmp1 and its recombinant expression vector and application
  • A Phytophthora inducible synthetic promoter pmp1 and its recombinant expression vector and application

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Experimental program
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Effect test

Embodiment 1

[0033] Example 1 Obtainment of a novel cis-element AAACTA

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

[0035] According to the microarray data literature of three different soybean varieties: V71-370, VPRIL9, and Sloan infected by Phytophthora sojae (Zhou L, Mideros SX, Bao L, et al. Infection and genotyperemodel 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 ne...

Embodiment 2

[0039] Example 2 The artificially synthesized PMP1 promoter is a Phytophthora inducible promoter

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

[0041] The new cis-element AAACTA obtained by bioinformatics analysis, and its upstream and downstream flanking sequences of 12 bp, a total of 30 bp, artificially synthesized four repeating units of these 30 bp, as the sequence of the artificially synthesized promoter PMP1 (such as SEQ ID NO.5 shown), used to construct plant expression vectors.

[0042] According to Chai C, Lin Y, Shen D, et al.Identification and FunctionalCharacterization of the Soybean GmaPPO12Promoter Conferring Phytophthorasojae Induced Expression[J].PloS One,2013,8(6):1-6.Construct the promoter fusion GUS report in the article The plant expression vector method of the gene obtains the PMP1::GUS recombinant plasmid that is fused with the GUS reporter gene by four repeating units. The construction pattern of the vector is as follows figure...

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Abstract

The invention belongs to the technical field of biology, and discloses a phytophthora-induced artificially-synthesized promoter PMP1 and a recombinant expression vector and application thereof. The promoter takes an AAACTA cis-element and 12 bp flanking sequences located at the upstream and downstream of the cis-element as a core sequence. A 30 bp sequence including the cis-element and 12 bp flanking sequences located at the upstream and downstream of the cis-element is taken, and is subjected to artificial synthesis to form a tetramer which is taken as the promoter PMP1. The PMP1 fused into a GUS reporter gene is constructed in a plant expression vector pMDC162, and through a tobacco transient expression system mediated by agrobacterium, the pathogenic-bacteria-introduced expression features of the promoter is analyzed. The result shows that the artificially-synthesized promoter PMP1 can fast and effectively drive up-regulation inducible expression of the GUS reporter gene through induction by phytophthora. It follows that the artificially-synthesized promoter PMP1 is a phytophthora-induced promoter, and therefore a valuable phytophthora-induced promoter is provided 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 PMP1 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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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/82C12N5/10A01H5/00A01H6/82
CPCC12N15/113C12N15/8222C12N15/8282C12N2310/10
Inventor 柴春月窦道龙曾文韬
Owner 江苏南京农大科技开发有限责任公司
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