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Promotor specificly-response to heavy metal ion

A technology for heavy metal ions and heavy metals, applied in the field of plant promoters, can solve problems such as not being ideal, and achieve the effects of reducing accumulation, improving resistance, and having great application value.

Inactive Publication Date: 2008-08-20
GRADUATE SCHOOL OF THE CHINESE ACAD OF SCI GSCAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Although the heavy metal resistance of model plants has been improved or the distribution of heavy metals in plants has been improved through plant genetic engineering technology, it is not very ideal overall, which has a great relationship with the selected promoter.
There is no relevant report on the study of the PvSR2 gene promoter

Method used

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  • Promotor specificly-response to heavy metal ion
  • Promotor specificly-response to heavy metal ion
  • Promotor specificly-response to heavy metal ion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1 Cultivation of plant material

[0025] Sterile tobacco seedlings (Nicotiana tabacum, W38) were grown in MS medium under 16h light (25°C) / 8h dark conditions. Arabidopsis thaliana (Columbiaecotype) seeds were surface sterilized and grown in 1 / 2 MS supplemented with 2% sucrose. The seeds of Phaseolus vulgaris L.cv. Saxa were sterilized and grown in soil pots. The growth conditions were 16h light (22°C) / 8h dark conditions. When the two first leaves are unfolded, use 0.2% (w / v) HgCl 2 . Spray vanes.

Embodiment 2

[0026] Embodiment 2. Bean PvSR2 gene promoter region clone

[0027] Step 1: DNA walking to obtain the upstream regulatory region of the gene

[0028] The genomic DNA of kidney bean was extracted by CTAB method, and the 5` upstream region of PvSR2 gene was cloned

[0029] Follow the instructions of the In vitro Cloning kit of Takara Bioengineering Co., Ltd. A brief description is as follows: the isolated bean genomic DNA was completely digested with PstI, HindIII and EcoRI three restriction endonucleases, recovered by ethanol precipitation, with the corresponding enzyme cleavage site and primers C1 and C1 provided in the kit. Connector connection for C2. Then use the gene-specific primer 1 (gene-specific primer1, GSP1, see Table 1) designed according to the PvSR2 gene and the primer C1 on the adapter (primer that comes with the kit) for the first amplification, and then dilute the amplification product Nested PCR was performed after 100 times, and the primers used were GSP2 ...

Embodiment 3

[0032] Example 3 5' Deletion Analysis Promoter Region Function

[0033] Step 1: Construction of promoter region-GUS reporter gene fusion expression vector

[0034] A fragment of a different region at the 5' end of the PvSR2 promoter region was linked with the GUS reporter gene to form a fusion gene. The details are as follows: primers (sense primers SP1, SP2, SP3, SP4, SP5 and SP6, antisense primers ASP) are designed according to the promoter region (see Table 1 for specific sequences), and the lengths of 1.6, 1.2, and 1.2 are obtained by PCR amplification respectively. 0.7, 0.3, 0.15 and 0.1kb promoter regions, the template used for PCR amplification is plasmid pUC-MRP, these promoter region deletion fragments correspond to figure 1 -1623 to +48, ​​-1161 to +48, ​​-687 to +48, ​​-281 to +48, ​​-146 to +48 and -90 to +48. HindIII and XbaI restriction sites were introduced at both ends in the PCR process of these fragments. These PCR products were digested with HindIII and Xb...

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Abstract

This invention is associated with the inducible promoter of new heavy metal in plant. This invention provides the promoter sequence of bean heavy metal inducive gene PvSR2 gene and the sequence of verified metalloid response element. Using the promoter in this invention to undergo the transgenic development of gene engineering products can improve the fastness to heavy metal of plant, reduce the accumulation of heavy metal in edible plant as well as increase the heavy metal accumulation capacity of transgenic plant and eukaryotic microbiology. This invention has significant utilization value in administer heavy metal pollution.

Description

Field of Invention [0001] The present invention belongs to the field of plant promoters, and in particular relates to a promoter responding to heavy metal ions in plants and a metalloid response element in the promoter sequence. Background technique [0002] Some heavy metals are essential trace elements for organisms, but excessive amounts can also cause harm, while non-essential heavy metals can cause damage to organisms. Using plant genetic engineering technology to develop transgenic plant products can improve the resistance of plants to heavy metals, reduce the toxicity of heavy metals in edible plants, and protect human health. Value. To achieve the above goals through plant genetic engineering, the influencing factors include the target gene of the protein or polypeptide that binds or transports heavy metal ions, the transgenic plant species, the transgenic method and the promoter used to control the expression of the target gene, among which the most critical is th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/29C12N15/82
Inventor 柴团耀祁晓廷张玉秀
Owner GRADUATE SCHOOL OF THE CHINESE ACAD OF SCI GSCAS