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Method and its expression carrier for eliminating environmental pollution from Tr-gene plant gene drift

A technology of transgenic plants and expression vectors, which is applied in the field of eliminating environmental pollution caused by gene drift of transgenic plants, and can solve problems such as unfavorable transgenic plant research and large-scale cultivation, no discovery, and damage to ecosystem biodiversity by foreign genes of transgenic plants

Inactive Publication Date: 2007-11-21
CHINA AGRI UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, the government and the public are worried about the possible environmental pollution caused by the genetic drift of transgenic plants, which is not conducive to the research and large-scale cultivation of transgenic plants (GMO Risk and Management (Edited by Xue Dayuan), 2005, China Environmental Science Press, Beijing, 280 pages)
2. The exogenous genes of transgenic plants may infiltrate wild relatives and destroy the original biodiversity of the ecosystem (Wei Wei et al., Acta Botanica, 1999, 41(4): 343-348)
Cytidine deaminase is found in bacteria and fungi, but it is not found in higher plants and mammalian cells (Stouggard J et al, Plant J.1993, 3: 755-761: Austin et al, Mol.Pharmacol ., 1993, 43: 380-387)

Method used

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  • Method and its expression carrier for eliminating environmental pollution from Tr-gene plant gene drift
  • Method and its expression carrier for eliminating environmental pollution from Tr-gene plant gene drift
  • Method and its expression carrier for eliminating environmental pollution from Tr-gene plant gene drift

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Embodiment 1, the construction of the plant expression vector p33UbdA containing chemically controlled bomb gene (codA gene) expression cassette and target gene (Bar gene) expression cassette expression cassette

[0058] According to the description of GenBank accession number AY331712, the codA gene with a length of 1284bp (sequence 2) was cloned from Escherichia coli JM101 genomic DNA, and restriction enzymes Bam HI and Sac I were added to its 5' end and 3' end respectively After the recognition site, it was ligated to the pGEM-T vector (Promega, USA) that had been double-digested with Bam HI and Sac I to generate the intermediate vector pdAJM for future use. The universal plasmid pAHC25 containing the GUS gene expression cassette driven by the Ubiquilin promoter (the structure and construction method of the plasmid is shown in: Chritensen AH et al., Plant Mol. Biol, 1992, 18: 675-689) was digested with Hind III, and recovered The fragment of ubi promoter-intron-GUS-n...

Embodiment 2

[0059] Embodiment 2, test tube obtains the transgenic plant of the plant expression vector p33UbdA containing chemically controlled bomb gene (codA gene) expression cassette and target gene (Bar gene) expression cassette

[0060] (1) Transformation of Agrobacterium

[0061] The plant expression vector p33UbdA constructed in Example 1 was transferred into the general-purpose Agrobacterium tumefaciens strain EHA105 (purchased from Dingguo Biotechnology company). Pick the single colony of transformed Agrobacterium grown on the YEB solid medium containing antibiotic rifampicin (Rif) 25mg / L, and culture it by shaking in YEB liquid medium containing antibiotic rifampicin (Rif) 25mg / L After 12 hours, plasmid DNA was extracted by the lye miniprep method (Sambrook et al., 2001, Handbook of Molecular Cloning). The plasmid was double digested with Bam HI and Sac I, and the 1.28kb (codA) target band and the vector band larger than 12kb were cut out to identify and confirm the success of...

Embodiment 3

[0071] Example 3. The test tube chemical detonation test of the plant expression vector p33UbdA transgenic plant containing chemically controlled bomb gene (codA gene) expression cassette and target gene (Bar gene) expression cassette

[0072] Get the blade of the transgenic tobacco test-tube plantlet of the target gene (Bar gene, anti-PPT gene) expression cassette and chemical control formula bomb gene (codA gene) expression cassette obtained by embodiment 2 and the blade of the non-transgenic control tobacco test-tube plantlet, cut into Discs with a diameter of 0.8-1 cm were inoculated into solid MS medium (MS+BA 2.0mg / L+IAA 0.5mg / L+inositol 100mg / L+sucrose 3%+agar powder 7g / L, pH5.8), cultured at 25±2°C under 16 light. After 30 days, it was observed that the transgenic leaf discs could not differentiate and shoot on the medium containing 100mg / L5-FC, but 65% (39 / 60) of the leaves had callus, while 80% (48 / 60) of the control leaves Discs can differentiate small buds. When ...

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Abstract

The invention provided a method to eliminate the gene shifting of the transforming gene plant and the expression carrier which includes the bomb gene expressing box and the target gene expressing box for avoiding the shift of the gene. The method includes converse the plant by the expressing carrier and get the conversion gene plant with the bomb gene expressing box which can express the target gene. Once the transforming plant is found to pollute the wild species, sub marginal species or the distant species, the gene bomb will be fired to kill the polluting plant. So the invention can solve the environment pollute by the transforming plant.

Description

technical field [0001] The invention relates to a method for eliminating environmental pollution caused by transgenic plant gene drift and its expression vector. Background technique [0002] The advent of the first transgenic plant (tobacco) in 1983 marked the beginning of a new round of "green revolution" in the world. After 20 years of research and development, the global planting area of ​​transgenic plants has exceeded 50 million hectares per year, and transgenic cotton, rapeseed, corn, soybeans and potatoes have entered the stage of large-scale commercial production. Since my country launched the research on transgenic plants in the middle and late 1980s, under the strong support of the national "863" plan, the special project of "Research and Industrialization of Transgenic Plants" and the Natural Science Foundation, the research and utilization of transgenic plants have made great progress. , the planting area of ​​transgenic plants exceeds 1.5 million hectares, beco...

Claims

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Application Information

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IPC IPC(8): C12N15/09C12N15/82C12N15/54
Inventor 肖兴国王志林曹克浩韩晓红孙晓红
Owner CHINA AGRI UNIV
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