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A Phytophthora inducible synthetic promoter pmp3 and its recombinant expression vector and application

An expression vector and a technology for inducing expression, applied in the biological field, can solve the problems of insignificant control effect, complicated control methods for Phytophthora disease, lack of pathogen-inducible promoters, etc.

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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  • 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 pmp3 and its recombinant expression vector and application
  • A Phytophthora inducible synthetic promoter pmp3 and its recombinant expression vector and application
  • A Phytophthora inducible synthetic promoter pmp3 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 novel cis-element AAAGAC

[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 new ...

Embodiment 2

[0039] Example 2 The artificially synthesized PMP3 promoter is a Phytophthora inducible promoter

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

[0041] The novel cis-element AAAGAC obtained by bioinformatics analysis, and its upstream 9bp and downstream 15bp flanking sequences, a total of 30bp, artificially synthesized four repeating units of these 30bp, as the sequence of the artificially synthesized promoter PMP3 (such as SEQID NO. 5), 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 PMP3::GUS recombinant plasmid fused with the GUS reporter gene by four repeating units, and the construction pattern of the vector is as follows figure 2 shown. ...

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Abstract

The invention belongs to the technical field of biology, and discloses a phytophthora inducible artificially synthesized promoter PMP3, as well as a recombinant expression vector and application thereof. The promoter adopts flanking sequences of an AAAGAC cis element, upstream 9bp and downstream 15bp thereof as core sequences. The cis element and flanking sequences of upstream 9bp and downstream 15bp of the cis element in a gene promoter, totally sequences of 30bp, are selected to artificially synthesize a tetramer sequence to serve as a promoter PMP3; the PMP3 fused GUS reporter gene is constructed into a plant expression vector pMDC162; and the pathogenic bacterium inducible expression characteristics of the promoter are analyzed by an agrobacterium induced tobacco transient expression system. Results discover that the artificially synthesized PMP3 promoter can be used for quickly and efficiently driving up-regulated inducible expression of the GUS reporter gene under induction of phytophthora. Thus, the artificially synthesized PMP3 promoter is a pathogenic inducible promoter, and a valuable pathogenic inducible promoter is provided to plant anti-phytophthora gene engineering.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a Phytophthora inducible artificially synthesized promoter PMP3 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
CPCC07K14/415C12N15/8239
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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