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Herbicide resistance protein, its encoding gene and use

A herbicide and protein technology, applied in genetic engineering, plant gene improvement, application, etc., can solve the problem of undiscovered 24DT16 herbicide resistance protein expression level, herbicide tolerance report, etc., to achieve increased flexibility and high flexibility sexual effect

Active Publication Date: 2016-04-20
BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There is no report on the expression level of 24DT16 herbicide resistance protein in plants and its tolerance to herbicides

Method used

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  • Herbicide resistance protein, its encoding gene and use
  • Herbicide resistance protein, its encoding gene and use
  • Herbicide resistance protein, its encoding gene and use

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example 、24D

[0093] The first embodiment, the acquisition and synthesis of 24DT16 gene sequence

[0094] 1. Obtain the 24DT16 gene sequence

[0095] The amino acid sequence (305 amino acids) of the 24DT16 herbicide resistance protein is shown in SEQ ID NO: 2 in the sequence listing; the amino acid sequence (305 amino acids) encoding the 24DT16 herbicide resistance protein corresponding to said 24DT16 herbicide resistance protein is obtained according to the plant preference codon ) nucleotide sequence (918 nucleotides), as shown in SEQ ID NO:1 in the sequence listing.

[0096] 2. Synthesize the above 24DT16 nucleotide sequence

[0097] The 24DT16 nucleotide sequence (shown as SEQIDNO: 1 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd.; the 5' end of the synthesized 24DT16 nucleotide sequence (SEQIDNO: 1) is also connected with SpeI restriction site, the 3' end of the 24DT16 nucleotide sequence (SEQ ID NO: 1) is also connected with a KasI restriction s...

no. 2 example

[0098] The second embodiment, construction of Arabidopsis recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0099] 1. Construction of Arabidopsis thaliana recombinant cloning vector DBN01-T containing 24DT16 nucleotide sequence

[0100] The synthesized 24DT16 nucleotide sequence was ligated into the cloning vector pGEM-T (Promega, Madison, USA, CAT: A3600), and the operation steps were carried out according to the instructions of the pGEM-T vector produced by Promega Company to obtain the recombinant cloning vector DBN01-T, which Build process such as figure 1 Shown (wherein, Amp represents the ampicillin resistance gene; f1 represents the replication origin of phage f1; LacZ is the LacZ start codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; 24DT16 is the 24DT16 nucleoside acid sequence (SEQ ID NO: 1); MCS is a multiple cloning site).

[0101] Then, the recombinant cloning vector DBN01-T...

no. 3 example

[0111] The third embodiment, the acquisition of Arabidopsis plants transferred to 24DT16 nucleotide sequence

[0112] Wild-type Arabidopsis seeds were suspended in 0.1% agarose solution. Suspended seeds were stored at 4°C for 2 days to fulfill the need for dormancy to ensure simultaneous seed germination. Mix horse manure with vermiculite and subirrigate with water until moist, allowing the soil mixture to drain for 24 hours. The pretreated seeds were planted on the soil mixture and covered with a moisture hood for 7 days. Let the seeds germinate and keep the light intensity at 120-150μmol / m at constant temperature (22°C) and constant humidity (40-50%) 2 Plants were grown in the greenhouse under long-day conditions (16 h light / 8 h dark) of 2 sec. Start by watering the plants with Hoagland's nutrient solution, followed by deionized water, keeping the soil moist but not soggy.

[0113] Transform Arabidopsis using the flower soak method. Inoculate one or more 15-30ml precult...

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Abstract

The invention relates to a weedicide-resistant protein, a coding gene and application thereof. The weedicide-resistant protein comprises: (a) protein composed of amino acid sequence disclosed as SEQ ID NO:2; or (b) (a)-derived protein with aryloxyanoate dioxygenase activity subjected to substitution and / or deletion and / or addition of one or more amino acids on the amino acid sequence in (a). The weedicide-resistant protein provided by the invention is especially suitable for expression in plants, and has wide resistance applicability to weedicides, especially phenoxy-auxin weedicides.

Description

technical field [0001] The present invention relates to a herbicide resistance protein, its encoding gene and application, in particular to a protein with resistance to 2,4-D, its encoding gene and application. Background technique [0002] Weeds can quickly deplete the soil of valuable nutrients needed by crops and other plants of interest. There are many types of herbicides currently used to control weeds, one particularly popular herbicide is glyphosate. Glyphosate-resistant crops have been developed, such as corn, soybean, cotton, sugar beet, wheat and rice. Fields of glyphosate-resistant crops can thus be sprayed with glyphosate to control weeds without significantly damaging the crop. [0003] Glyphosate has been widely used globally for more than 20 years, resulting in an overreliance on glyphosate and glyphosate-tolerant crop technologies, and in wild weed species that are naturally more tolerant to glyphosate or have developed Plants exhibiting glyphosate resista...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/02C12N15/53C12N15/63C12N15/84C12N5/10C12N1/13C12N1/19C12N1/21C12N7/01A01K67/033A01H5/00A01G13/00
CPCC12N9/0071C12N15/8209C12N15/8274
Inventor 吴业春鲍晓明牛晓广康越景丁德荣
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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