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Promoter for regulating expression of genes in T shape trichome and application of promoter

A non-secretory glandular trichome and gene regulation technology, applied in the field of promoters, can solve the problems of temporal and spatial regulation of target genes, excessive consumption of cellular material and energy, and harm

Active Publication Date: 2015-11-25
SHANGHAI JIAO TONG UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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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 expression of genes in T shape trichome and application of promoter
  • Promoter for regulating expression of genes in T shape trichome and application of promoter
  • Promoter for regulating expression of genes in T shape trichome and application of promoter

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Embodiment

[0038] This embodiment relates to the acquisition of the AaTTPS5 gene promoter, which specifically includes the following steps:

[0039] Step 1. Cultivate aseptic seedlings of Artemisia annua

[0040] 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 MS solid medium without any hormone, 25°C, 16h / 8h (light / dark) light culture, after 14 days, you can obtain the sterile vaccine of Artemisia annua;

[0041] Step 2: Cloning of the promoter sequence in the genomic DNA

[0042] 1. Genomic DNA Extraction

[0043] Add 2 steel balls into a 1.5mL centrifuge tube, and put a piece of Artemisia annua leaf (1cm 2 About the size, take it in an ice box). Add 300 μL TPS buffer (operate in a fume hood, TPS contains 2% mercaptoethanol), shake at 55-60 Hz for 90 seconds. Then add 300 ...

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Abstract

The invention discloses a gene promoter, belonging to the field of plant biotechnology. The nucleotide sequence of the promoter is as shown in SEQ ID NO. 1, and the promoter is capable of regulating the specific expression of target genes in T shape trichome of a plant. Meanwhile, the invention further relates to application of the gene promoter in genetic engineering breeding which realizes expression of a gene in trichome tissues of the plant and produces metabolites. When the promoter realizing specific expression of the gene in trichome of the plant is used for carrying out genetic manipulation on the trichome system of the plant, no harm is caused to the growth or development of the plant; therefore, the promoter disclosed by the invention has an important significance for the genetic engineering breeding which realizes expression of the gene in trichome tissues of the plant and produces metabolites.

Description

technical field [0001] The present invention relates to a promoter, in particular to a terpene synthase gene promoter (proTTPS5) obtained by means of molecular biology and specifically expressed in plant non-secretory glandular trichomes and its application in transgenic plants . Background technique [0002] Artemisia annua L. is an annual herb of the genus Artemisia in the family Asteraceae. The plant has a strong volatile aroma, and its plant contains many secondary metabolites: artemisinin, volatile oil, α-pinene, camphor, artemisinone, etc. , in addition to containing flavonoids. Artemisinin (Artemisinin) is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is recommended as an antimalarial drug combination therapy (Artemisinin-based combination therapies, ACTs) main active ingredients. At present, the main source of artemisinin is extracted from the aerial parts of Artemisia annua, however, the content of artem...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/82C12Q1/68A01H5/00
Inventor 唐克轩石璞付雪晴沈乾孙小芬江伟民吕宗友颜廷祥
Owner SHANGHAI JIAO TONG UNIV
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