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A kind of aapdr2 gene promoter and its function verification method and application

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: 2018-09-11
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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  • A kind of aapdr2 gene promoter and its function verification method and application
  • A kind of aapdr2 gene promoter and its function verification method and application
  • A kind of aapdr2 gene promoter and its function verification method and application

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Embodiment

[0045] The present embodiment relates to the acquisition and functional verification of the AaPDR2 gene promoter, which specifically includes 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, and dried with sterile absorbent paper. On hormone-free MS solid medium, 25 ℃, 16h / 8h (light / dark) light culture, 14 days later, the sterile seedlings of Artemisia annua were obtained;

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

[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 based on the The specific primers designed for 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 a method for verifying its function and application. Background technique [0002] Artemisia annua L. is an annual herb of the genus Asteraceae. Artemisinin, a sesquiterpene lactone oxide containing a peroxy bridge extracted from its aerial parts, is currently the most widely used and most effective antimalarial drug, especially for cerebral malaria and anti-chloroquine malaria. . Currently, artemisinin combination therapy (ACTs) is 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 trichomes on the leaf surface of Artemisia annua: secretory glandular trichome (GST) and non-secretory glandular trichome (T shape trichome, TST). Transmission and so on all play a cru...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/84C12Q1/6897A01H5/00
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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