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DXR promoter for lily flower part peculiarities and wound inductions and application of DXR promoter

A technology of injury induction and promoter, applied in the field of genetic engineering, can solve the problems of no time-space specificity, affect the survival rate of positive plants, affect the normal growth and development of plants, etc., and achieve the effect of shortening the breeding time

Inactive Publication Date: 2015-11-11
SOUTH CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, the choice of promoters in transgenic technology mainly adopts constitutive promoters. Under the control of constitutive expression promoters, foreign genes can be expressed in all parts of transgenic plants and at different developmental stages, and can be expressed continuously and efficiently. No space-time specificity, resulting in energy loss, affecting the survival rate of positive plants, causing abnormal changes in plant physiology and morphology, thus affecting the normal growth and development of plants

Method used

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  • DXR promoter for lily flower part peculiarities and wound inductions and application of DXR promoter
  • DXR promoter for lily flower part peculiarities and wound inductions and application of DXR promoter
  • DXR promoter for lily flower part peculiarities and wound inductions and application of DXR promoter

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0034] Obtaining of DXR promoter sequence

[0035] S1. Extraction of total DNA from lily petals: 0.1-0.2 g of lily plant petal powder ground with liquid nitrogen was transferred to a 2.0 mL centrifuge tube. Add 1 mL of preheated 2% CTAB solution, mix thoroughly, and bathe in 65°C water for 30 minutes; every 10 minutes, shake upside down once. Centrifuge at 12,000rpm for 5 minutes, transfer the supernatant to a new centrifuge tube, then add an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1) to the centrifuge tube, invert several times, shake gently until the solution becomes emulsified , 12,000rpm, centrifuge for 5min, transfer the supernatant to a new centrifuge tube. Repeatedly add an equal volume of phenol / chloroform / isoamyl alcohol (25:24:1) once, centrifuge at 12,000rpm for 5min. Take the supernatant, add 2 / 3 volume of isopropanol, and place it at -20°C for 30min. Add 1×TE solution to dissolve the precipitate, add 1 / 10 volume of NaAC (3mol / L) and 2 volumes o...

Embodiment 2

[0049] Fusion Expression of DXR Promoter and GUS in Transformed Tobacco

[0050] S1. The sequence of the DXR promoter obtained according to Example 1, designed to include XbaI and HindIII Primers used to amplify the three 5'-terminal deletion sequences p-534, p-767 and p-969 at restriction sites. The primer sequences are as follows:

[0051] Primers for amplifying p-534:

[0052] p-534-F: 5'-GGCAAGCTTACCATACATGTGACACACAG-3' (SEQ ID NO: 14)

[0053] Primers for amplifying p-767:

[0054] p-767-F: 5'-GGCAAGCTTCTGACCCGTTTATAAACGAG-3' (SEQ ID NO: 15)

[0055] Primers for amplifying p-969

[0056] p-969-F: 5'-GGCAAGCTTCCTCACAAGATAGACGATC-3' (SEQ ID NO: 16)

[0057] Downstream primers are:

[0058] R: 5'-GGGCCTCTAGATTGAGGAGAGAGAAAGAGATAAG-3' (SEQ ID NO: 17)

[0059] S2.pCAMBIA1300G plant expression vector XbaI and HindIII Restriction endonucleases were used for double digestion, and large fragments were recovered on 1% agarose gel. Directly insert the target fragment int...

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Abstract

The invention belongs to the technical field of gene engineering and particularly discloses a DXR promoter for lily flower part peculiarities and wound inductions and application of the DXR promoter. The nucleotide sequence of the promoter is shown in SEQ ID NO:2. The target gene regulated by the DXR promoter is only expressed in the flower tissue in transgenic tobacco, and the expression of the target gene is related to the flower development process and is induced by damage and jasmonic acid methyl ester. In addition, the DXR promoter can be connected to any plant transformation carrier and guided into lilies or other plant cells, the transgene flowery flavor containing the promoter and resistance can be obtained, and therefore the promoter can be used for production. Peculiar molecular markers can also be generated according to promoter sequence information, used for identifying lilies or other plant flowery flavors and resistance gene types and used for molecular marker auxiliary selective breeding, and therefore the selecting efficiency of breeding is improved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a lily flower-specific and injury-inducible DXR promoter and application thereof. Background technique [0002] The promoter is a DNA sequence located in the upstream region of the 5' end of the structural gene that can recognize and activate RNA polymerase to accurately combine with the template DNA to ensure accurate and efficient initiation of transcription. It is the center of gene transcription regulation in plants, and is the key to understanding gene transcription regulation mechanism and expression pattern. [0003] According to the transcription mode of promoters, they can be divided into: constitutive promoters, tissue or organ-specific promoters and inducible promoters. Among them, tissue-specific promoters are also called organ-specific promoters, including fruit-specific promoters, root-specific promoters, leaf-specific promoters, and floral organ-specif...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/63C12N1/21C12N5/10C12N15/82A01H5/00
Inventor 范燕萍李昕悦余让才吴萌萌玉云祎岳跃冲
Owner SOUTH CHINA AGRI UNIV
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