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AaPDR2 gene promoter as well as functional verification method and application thereof

A technology for functional verification and promoter, applied in application, genetic engineering, plant genetic improvement, etc., can solve the problem that the promoter sequence of AaPDR2 gene has not yet been found.

Active Publication Date: 2016-07-20
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

After searching the prior art literature, it is found that there is no report related to the AaPDR2 gene promoter sequence of the present invention

Method used

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  • AaPDR2 gene promoter as well as functional verification method and application thereof
  • AaPDR2 gene promoter as well as functional verification method and application thereof
  • AaPDR2 gene promoter as well as functional verification method and application thereof

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Embodiment

[0045] This embodiment involves the acquisition and functional verification of the AaPDR2 gene promoter, specifically including the following steps:

[0046] Step 1. Cultivate aseptic seedlings of Artemisia annua

[0047] 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 a hormone-free MS solid medium at 25°C under 16h / 8h (light / dark) light, and after 14 days, you can obtain an Artemisia annua sterile vaccine;

[0048] Step 2, according to the AaPDR2 gene promoter sequence in the whole genome of Artemisia annua, clone the promoter sequence

[0049] In order to improve the specificity of the product, two rounds of nested PCR amplification were used, and the primers used were shown in Table 2. AaPDR2-FP1 (SEQ ID NO.2) and AaPDR2-RP1 (SEQ ID NO.3) were obtained according t...

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Abstract

The invention relates to an AaPDR2 gene promoter as well as a functional verification method and application thereof. The AaPDR2 gene promoter comprises the following cis-acting elements: ABRE, BoxI, CAAT-box, CCAAT-box, CGTCA-motif, G-Box, G-box, GA-motif and TATA-box. Further, the DNA (Deoxyribonucleic Acid) sequence of the AaPDR2 gene promoter is shown in SEQ ID NO.1. The AaPDR2 gene promoter has a function of predominantly expressing genes guided by the AaPDR2 gene promoter in glandular secretory trichome and T shape trichome of young leaves of Artemisia annua L., and the function is confirmed by a GUS reporter gene. Therefore, the AaPDR2 gene promoter disclosed by the invention has important significance for genetic engineering breeding for expressing and generating metabolites by using plant glandular trichome tissues.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an AaPDR2 gene promoter and its function verification method and application. Background technique [0002] Artemisia annua L. is an annual herb belonging to the family Asteraceae. Artemisinin, a sesquiterpene lactone oxide containing a peroxy bridge extracted from its aerial part, is currently the most widely used and effective antimalarial drug, especially for cerebral malaria and anti-chloroquine malaria. . Currently, artemisinin combination therapies (ACTs) are the most effective treatment for malaria recommended by the World Health Organization. However, the content of artemisinin in the plant Artemisia annua is low, which cannot fully meet the global market demand. 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). It is also cl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12N15/84C12Q1/68A01H5/00
CPCC07K14/415C12N15/8225C12Q1/6897
Inventor 唐克轩何倩刘萌石璞付雪晴郝小龙黎凌孙小芬
Owner SHANGHAI JIAO TONG UNIV
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