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Promoter for regulating and controlling expression of genes in non-secretory type glandular hairs and application of promoter

A non-secretory glandular hair, gene regulation technology, applied in the field of plant biology, can solve the problems of time and space regulation of target genes, excessive consumption of cell material and energy, harm and other problems

Active Publication Date: 2015-12-23
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current genetic engineering research of Artemisia annua, constitutive promoters are often used, such as the cauliflower mosaic virus 35S promoter (CaMV35S), which can drive the expression of foreign genes in all tissues and organs, and will cause excessive consumption. The substance and energy in the cell, and the expression of the target gene cannot be regulated in time and space, which is likely to cause a certain burden and harm to the normal growth of plants

Method used

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  • Promoter for regulating and controlling expression of genes in non-secretory type glandular hairs and application of promoter
  • Promoter for regulating and controlling expression of genes in non-secretory type glandular hairs and application of promoter
  • Promoter for regulating and controlling expression of genes in non-secretory type glandular hairs and application of promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] Cloning of the Genome Sequence of the Promoter from the Artemisia annua Genome

[0031] 1. Cultivate aseptic vaccines of Artemisia annua:

[0032] Artemisia annua seeds were soaked in ethanol with a volume fraction of 75% for 1 min, then soaked in 20% (w / v) NaClO for 20 min, rinsed with sterile water for 3 to 4 times, blotted the surface moisture with sterile absorbent paper, and inoculated Cultivate on hormone-free MS solid medium at 25°C and 16h / 8h (sunlight / darkness) under light, after 14 days you can obtain sterile A. annua seedlings;

[0033] 2. Genomic DNA extraction

[0034] Put a leaf of Artemisia annua (1cm 2 About the size, take it in an ice box), add 2 steel balls. Add 300 μL TPS buffer (operate in a fume hood, TPS contains mercaptoethanol), shake at 55-60 Hz for 100 seconds. Then add 300 μL TPSbuffer (fume hood). Water bath at 65°C for 1h (shake every 20min), and the time can be extended appropriately, up to 1.5h. Cool to room temperature and centrifug...

Embodiment 2

[0050] Analyze the cis-acting elements of the AaTPS7 gene promoter to determine the type of AaTPS7 gene promoter

[0051] The length of the AaTPS7 gene promoter sequence obtained in the present invention is 1167bp. In order to find the cis-acting elements on the promoter, the promoter of AaTPS7 gene was analyzed with Plantcare (http: / / bioinformatics.psb.ugent.be / webtools / plantcare / html / ). The analysis found that in addition to TATAbox and CAATbox, there are many cis-acting elements on the promoter of polycloning: CGTCA-motif, HSE, MBS, MRE, TC-richrepeats, WUN-motif, etc.

[0052] The cis-acting elements in the promoter sequence are shown in Table 4:

[0053] Table 4: Analysis of regulatory elements in the promoter sequence

[0054]

[0055] In Table 4, the positions of -980, -907, -678 and -130 refer to the corresponding positions in the sequence listed in Example 1.

[0056] CGTCA-motif is a methyl jasmonate response element; HES responds to heat stress; MBS and MRE ar...

Embodiment 3

[0058] Connect the obtained promoter into the pCAMBIA-1391z vector and fuse the GUS reporter gene

[0059] In order to study the expression of the gene promoter in different plant parts, the promoter proTPS7 of the AaTPS7 gene was connected to the pCAMBIA-1391z vector and fused with the GUS reporter gene. site, the NcoI restriction site was introduced into the reverse primer, and the primer sequence is shown in the table below:

[0060] Table 5 pCAMBIA1391z-proTPS7 vector constructs PCR primers

[0061]

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Abstract

The invention discloses a promoter for regulating and controlling expression of genes in non-secretory type glandular hairs. The nucleotide sequence of the promoter is shown in SEQ ID No:1. The invention further provides a preparation method of the promoter, a carrier containing the promoter and an expression case. The promoter can regulate and control the expression of target genes in time and space, therefore, morphogenesis and development of the non-secretory type glandular hairs and metabolism regulation of secondary metabolites in the non-secretory type glandular hairs can be intensively studied, and application in genetic engineering breeding for producing the metabolites can be achieved. Therefore, the promoter for regulating and controlling the expression of the genes in the non-secretory type glandular hairs has important significance on genetic engineering breeding for expressing and producing the metabolites by means of plant glandular hair tissue.

Description

technical field [0001] The invention relates to the field of plant biotechnology, in particular to a terpene synthase gene promoter (proTPS7) specifically expressed in plant non-secretory glandular hairs and its application in transgenic plants. Background technique [0002] Artemisia annua (Artemisia annua L.) is an annual herb of the genus Artemisia in the family Compositae. The plant has a strong volatile aroma. The essential oil of Artemisia annua extracted from the glandular hairs of Artemisia annua contains a large number of terpenoids and flavonoids with pharmacological or biocidal activity. class of compounds. There are two kinds of glandular hairs on the leaf surface of Artemisia annua: secretory glandular hairs (glandular secretory trichome, GST) and non-secretory glandular hairs (Tshape trichome, TST). to a vital role. Artemisinin is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is currently the most ef...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/84A01H5/00
Inventor 唐克轩陈俏沈乾付雪晴石璞孙小芬颜廷祥
Owner SHANGHAI JIAO TONG UNIV
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