Artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence

A technology of cytidine diphosphate and coding sequence, which is applied in the field of coding sequence and coding sequence of Artemisia annua 4--2-C-methyl-D-erythritol synthase, and can solve problems such as no coding sequence reports yet, Achieve the effect of realizing large-scale production and increasing content

Inactive Publication Date: 2010-03-10
SUZHOU TANGJI BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] After searching the literature of the prior art, it is found that there is no report related t

Method used

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Embodiment

[0023] Step 1, cloning of Artemisia annua 4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence (CMS)

[0024] (1) RNA extraction (RNA extraction)

[0025] Take the leaf tissue of Artemisia annua, grind it in liquid nitrogen, add it to a 1.5mL Eppendorf (EP) centrifuge tube filled with lysate, shake it fully, and extract total RNA according to the instructions of the TIANGEN kit. The quality of total RNA was identified by formaldehyde denaturing gel electrophoresis, and then the RNA content was determined on a spectrophotometer.

[0026] (2) Full-length cloning of the gene (Cloning of Full-length cDNA)

[0027] According to the conserved sequence of amino acids encoded by related genes in Arabidopsis, using the principle of homologous gene cloning, the RACE (Rapid Amplification of cDNA Ends) method (SMART of Clontech Company) was adopted. TM RACE kit) for full-length cDNA cloning in three stages:

[0028] ①Synthesis of first-strand cDNA

[0029] Take...

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PUM

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Abstract

The invention relates to an artemisia apiacea4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase coding sequence in the technical field of gene engineering. The sequence is a sequence generated after one or a plurality of codons in nucleotide are replaced by degenerate codons of amino acid with same coding, as shown in 18-position to 926-position of SEQ ID NO: 3 or shown in 18-position to926-position of SEQ ID NO: 3. The method for improving the content of plant artemisinin by using the coding sequence includes the steps of: connecting the coding sequence to a plant expression regulatory sequence and constructing a plant expression carrier containing the coding sequence; transferring the expression carrier in step 1 to agrobacterium rhizogenes, then transferring agrobacterium rhizogenes to artemisia apiacea; through antibiotic screening, obtaining a transformation cell containing the coding sequence and regenerating a transgenosis plant. The invention relates to a polypeptide, and the sequence is shown in SEQ ID NO: 4. The coding sequence can improve the content of artemisinin in artemisia apiacea and the content of isoprene compound of other plants.

Description

technical field [0001] The present invention relates to a coding sequence in the technical field of genetic engineering, in particular to a coding sequence of Artemisia annua 4-(5'-cytidine diphosphate)-2-C-methyl-D-erythritol synthase. Background technique [0002] Artemisia annua L. is an annual herb belonging to the family Asteraceae. Artemisinin is a sesquiterpene lactone compound containing a peroxide bridge structure isolated from its aerial part. It is currently recognized as the most effective drug for treating malaria in the world, especially for cerebral malaria and chloroquine-resistant malaria. efficient. Currently, the most effective treatment for malaria recommended by the World Health Organization is artemisinin combination therapy (ACTs). In addition, with the gradual deepening of pharmacological research on artemisinin, scientists have found that artemisinin and its derivatives also have anti-inflammatory, anti-schistosomiasis, anti-tumor and immune regula...

Claims

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Application Information

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IPC IPC(8): C12N15/52C12N9/00C12N15/82C12N15/84
Inventor 唐克轩王玥月杨可翁纯纯
Owner SUZHOU TANGJI BIOTECH CO LTD
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