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Selenium bioconversion relevant nucleotide sequence and application

A nucleotide sequence and biotransformation technology, applied in the field of nucleotide sequences related to selenium biotransformation, can solve the problems of low utilization rate of selenium element biotransformation, selenium flowing into farmland, and high cost, so as to improve human immunity and avoid pollution. Soil, the effect of reducing planting costs

Inactive Publication Date: 2020-08-25
张家港普渡中药饮片有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some of these selenium-enriched areas are small in size, and some areas are accompanied by undesirable minerals such as lead and mercury.
[0005] On the one hand, the area of ​​selenium-enriched areas is small, and the output is not enough to supply demand. On the other hand, due to the low biotransformation and utilization rate of selenium in rice, the method of spraying selenium is expensive, and a large amount of selenium spraying will cause unsatisfactory production. Selenium transformed and utilized by plants flows into farmland, causing water and soil pollution

Method used

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  • Selenium bioconversion relevant nucleotide sequence and application
  • Selenium bioconversion relevant nucleotide sequence and application
  • Selenium bioconversion relevant nucleotide sequence and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Take stratified sampling (two layers) and five-point sampling method, in selenium-enriched area (Guizhou Liuzhi Changtian village) in the soil sampling of general planting depth, sampling depth is 6cm and 12cm, every point sampling 100g, totally sampling 1000g, mixing Uniform detection.

[0022] Take 50g of the above-mentioned mixed sample and 100mL of buffer solution and mix them into a dialysis bag, dialyze for 12 hours to remove the salt ions in the sample, use the freeze-thaw method to lyse the cells and extract the total microbial DNA, purify the DNA sample, sequence the purified DNA and use the large intestine The bacillus expression system (E.coli DH5a) constructs the microbial gene library in the selenium-rich soil, and randomly breaks the DNA of the successfully constructed gene library into fragments of 100bp, 150bp, 200bp, 250bp, 500bp, 1000bp, 1500bp, and 2500bp.

[0023] For small fragments of DNA, connect the small fragments of DNA (100bp, 150bp, 200bp, 25...

Embodiment 2

[0026] Carry out codon modification to the Gfk3 nucleotide sequence, Gfk7 nucleotide sequence, and Gfk8 nucleotide sequence to adapt to the plant coding system, and obtain the modified nucleotide sequence O-Gfk3 (as shown in SEQ ID NO.1) , O-Gfk7 (as shown in SEQ ID NO.2), O-Gfk8 (as shown in SEQ ID NO.3). Sequence synthesis entrusted Nanjing GenScript Company to connect the synthesized O-Gfk3 nucleotide sequence, O-Gfk7 nucleotide sequence, and O-Gfk8 nucleotide sequence into the cloning vector pGEM-T, and the operation steps followed the pGEM product of Promega Company -T vector instructions are carried out.

[0027] Recombinant cloning vector Gzg-1103, recombinant cloning vector Gzg-1107, and recombinant cloning vector Gzg-1108 were obtained respectively. The structure of the recombinant cloning vector Gzg-1103 is ampicillin resistance gene, origin of replication of phage f1, LacZ initiation codon, SP6 RNA polymerase promoter, T7 RNA polymerase promoter, O-Gfk3 nucleotide ...

Embodiment 3

[0032] According to the method for constructing recombinant cloning vectors Gzg-1103, Gzg-1107, and Gzg-1108 described above, the recombinant cloning vectors Gzg-1103, Gzg-1107, and Gzg-1108 and the expression vector pCAMBIA1200 were digested with restriction endonucleases NcoI and HindIII, respectively. , insert the excised O-Gfk3, O-Gfk7, and O-Gfk8 nucleotide sequence fragments into the restriction sites of the corresponding expression vectors, and use conventional restriction methods to construct vectors is well known to those skilled in the art , constructed into recombinant expression vector Gzg-110003 (kanamycin gene; right border; promoter (SEQ ID NO:4); O-Gfk3 nucleotide sequence (SEQ ID NO:1); terminator (SEQ ID NO :5); LB: left border); Gzg-110007 (kanamycin gene; right border; promoter (SEQ ID NO:4); O-Gfk7 nucleotide sequence (SEQ ID NO:2); terminator (SEQ ID NO:5); LB: left border); Gzg-110008 (kanamycin gene; right border; promoter (SEQ ID NO:4); O-Gfk8 nucleoti...

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Abstract

The invention relates to a selenium bioconversion relevant nucleotide sequence and an application. The selenium bioconversion relevant nucleotide sequence comprises a nucleotide sequence as shown in SEQ ID NO.2 or SEQ ID NO.3. The invention also provides a method for planting selenium-enriched plants. The method comprises the step of introducing the selenium bioconversion relevant nucleotide sequence, or a recombinant clone carrier or a recombinant expression carrier to a plant genome. Selenium-enriched rice provided by the invention is not limited in planting lands, and is suitable for various farmland environments, and the purpose of enriching selenium elements can also be achieved even in planting environment having a general selenium content.

Description

technical field [0001] The invention relates to the field of plant production, in particular to a nucleotide sequence related to selenium biotransformation. Background technique [0002] Selenium is a rare and dispersed semi-metallic chemical element between metals and non-metals. The element symbol is Se. In 1973, the World Health Organization announced that selenium is an essential trace element for human body. Selenium belongs to the oxygen family element, and the selenium content in the natural crust is very small. Selenium is divided into inorganic selenium and organic selenium. Inorganic selenium is contained in minerals and is insoluble in water. It is not easy for the human body to accept and has toxic side effects. Organic selenium just makes up for the deficiency of inorganic selenium. There are more than 40 kinds of human diseases related to selenium deficiency, such as cardiovascular disease, cancer, peptic ulcer, diabetes, hepatitis, myopia, cataract, male dyst...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/70C12N15/82A01H5/10A01H6/46
CPCC07K14/195C07K14/37C12N15/70C12N15/8243
Inventor 陆开峰
Owner 张家港普渡中药饮片有限公司
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