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Artemisia annua absorbing formaldehyde and its cultivation method

A technology for absorbing formaldehyde and Artemisia annua, applied in the field of genetic engineering to cultivate plants, can solve the problem of no household plants, etc., and achieve the effect of improving the ability to absorb formaldehyde

Inactive Publication Date: 2018-01-12
CHONGQING MEDICAL & PHARMA COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] However, there are currently no home plants that can be directly used for formaldehyde absorption in home decoration.

Method used

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  • Artemisia annua absorbing formaldehyde and its cultivation method
  • Artemisia annua absorbing formaldehyde and its cultivation method
  • Artemisia annua absorbing formaldehyde and its cultivation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] 1.1 Shmt and Fdm gene PCR amplification and prokaryotic expression vector construction and enzyme protein expression cloning Shmt degenerate primers:

[0044] Upstream primer Primer1: 5'-ATGCGAATTTTACCGGGTTTTT-3', as shown in SEQ ID NO:3.

[0045] Downstream primer Primer2: 5'-TCGGCTGACATAATAAAACCAACGG-3', as shown in SEQ ID NO:4.

[0046] Clone Shmt PCR primers were designed according to its conserved amino acid sequence:

[0047] Upstream primer Primer3: 5′-AG CATATG CTGGCTAGCTGTTAGCGTTTT-3' (the underlined part is the NdeI restriction site), as shown in SEQ ID NO:5.

[0048] Downstream primer Primer6: 5′-CG CTCGAGG GGCTATAAGTAAACTGAAACGG-3′, (the underlined part is the XhoI restriction site), as shown in SEQ ID NO:6.

[0049] The genomic DNA of Pseudomonas putida was extracted according to the Molecular Cloning Guide, and the PCR reaction conditions were as follows: pre-denaturation at 97°C for 5 minutes, denaturation at 92°C for 1 minute, annealing at 56°C for...

Embodiment 2

[0067] Example 2 Introducing the Shmt and Fdm genes contained in the recombinant plasmid pCB-Fdm-Shmt into the Artemisia annua genome

[0068] 1) The pCB-Fdm-Shmt plasmid prepared above was transformed into Agrobacterium competent cell LBA4404 by electric shock method: pCB-Fdm-Shmt was introduced into Agrobacterium competent cell LBA4404 to obtain LBA4404 / pFdm-Shmt. The implementation steps are based on the English photocopy version 5 "Molecular Biology Mustard Experiment Manual" (Science Press. 2002), using the Agrobaterium (Agrobacterium) electroporation program that comes with the electroporation instrument of Biorad Company. The Agrobacterium into which the plant expression vector was introduced was used for the transformation of Artemisia annua.

[0069] 2) Transformation and regeneration of Artemisia annua: first, transform Agrobacterium, prepare Agrobacterium competent and transform, apply the transformed Agrobacterium liquid to a medium containing kanamycin 60Lg / mL, ri...

Embodiment 3

[0071] Example 3 Using real-time quantitative PCR to analyze the expression levels of Shmt and Fdm genes in positive Artemisia annua plants

[0072] Total RNA was extracted from the bioengineered Artemisia annua plants that were positive by PCR, and reverse-transcribed into cDNA.

[0073] Design primers for the internal reference gene β-actin, and calibrate the internal reference.

[0074] Upstream primer: 5'-GCCACCGATCCAGACACTGTACTTCC-3' as shown in SEQ ID NO:11

[0075] Downstream primer: 5'-ATTCAGGTGATGGTGTGAGCCACAC-3' as shown in SEQ ID NO:12

[0076] Target gene Shmt target gene detection primers (detection primers are shown in SEQ ID NO: 5, 6) as before. Amplification conditions: pre-denaturation at 94°C for 4 min; denaturation at 92°C for 30 S, annealing at 60°C (internal reference gene) or 66°C (adjustable target gene) for 30 S, 38 cycles, extension at 72°C for 10 min, and draw the curve. Such as Figure 4 It can be seen that the expression level of the target gene...

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Abstract

The invention provides artemisia apiacea absorbing formaldehyde and a cultivation method of the artemisia apiacea. The artemisia apiacea absorbing formaldehyde contains a Shmt gene and an Fdm gene. The sequence of the Shmt gene is SEQ ID NO: 1 and the sequence of the Fdm gene is SEQ ID NO: 2. According to the invention, a serine hydroxymethyltransferase gene Shmt of pseudomonas putida and the Fdm gene of a formaldehyde dismutase gene are cloned and converted into artemisia apiacea so as to improve synthesis of serine hydroxymethyltransferase and formaldehyde dismutase in artemisia apiacea, so that a genetic engineering artemisia apiacea variety with high content of serine hydroxymethyltransferase and formaldehyde dismutase is obtained, thereby greatly improving the capacity of artemisia apiacea absorbing formaldehyde. Therefore, the artemisia apiacea becomes a plant of eliminating formaldehyde released in home decoration.

Description

technical field [0001] The invention relates to the field of cultivating plants by using genetic engineering, in particular to cultivating an Artemisia annua capable of absorbing formaldehyde by using transgenic technology. Background technique [0002] Indoor air pollution mainly comes from furniture, decoration, smoking, kitchen fumes, gas water heaters, etc., which can be divided into four categories. The first category is volatile organic compounds, such as formaldehyde, benzene, ammonia and other chemical substances. NH 3 , CO, SO 2 Inorganic compounds such as carbon dioxide, as well as smoke from fuel combustion can be inhaled particulate matter. The second type of radioactive substance pollution such as radon, such as from construction slag bricks, cement, cement radioactive pollutants and decorative stone; the third type of microbial pollution, mainly from bacteria, fungi, virus spores, etc. from dogs, cats, cockroaches, bedding and toys, etc. The fourth category i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/84A01H5/00
Inventor 陈俊意朱照静杨治国杨延音谭丽田数高管琴张宝勇彭坤曾祥琼
Owner CHONGQING MEDICAL & PHARMA COLLEGE
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