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Herbicide resistance protein, its encoding gene and use

A resistance gene and herbicide technology, applied in genetic engineering, plant gene improvement, application, etc., can solve the problem of undiscovered 24DT21 herbicide resistance protein expression level, herbicide tolerance report, etc., to improve flexibility, The effect of high flexibility

Active Publication Date: 2019-10-18
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 24DT21 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

[0097] The first embodiment, the acquisition and synthesis of 24DT21 gene sequence

[0098] 1. Obtain the 24DT21 gene sequence

[0099] The amino acid sequence (292 amino acids) of the 24DT21 herbicide resistance protein, as shown in SEQ ID NO: 2 in the sequence listing; obtain the amino acid sequence (292 amino acids) corresponding to the 24DT21 herbicide resistance protein encoded according to the plant preference codon amino acids) nucleotide sequence (879 nucleotides), as shown in SEQ ID NO:1 in the sequence listing.

[0100] 2. Synthesize the above-mentioned 24DT21 nucleotide sequence

[0101] The 24DT21 nucleotide sequence (shown as SEQ ID NO:1 in the sequence listing) was synthesized by Nanjing KingScript Biotechnology Co., Ltd.; the 5' of the synthesized 24DT21 nucleotide sequence (SEQ ID NO:1) The end is also connected with a SpeI restriction site, and the 3' end of the 24DT21 nucleotide sequence (SEQ ID NO: 1) is also connected with a KasI restriction site.

no. 2 example

[0102] The second embodiment, construction of Arabidopsis thaliana and soybean recombinant expression vector

[0103] 1. Construction of recombinant cloning vector DBN01-T containing 24DT21 nucleotide sequence

[0104] The synthetic 24DT21 nucleotide sequence was connected 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 of the product of 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 initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; 24DT21 is the 24DT21 nucleoside acid sequence (SEQ ID NO: 1); MCS is the multiple cloning site).

[0105]Then, the recombinant cloning vector DBN01-T was transformed into Escherichia coli T1 competent cells (...

no. 3 example

[0113] The third embodiment, the acquisition of Arabidopsis plants transferred to 24DT21 nucleotide sequence

[0114] 1. Transformation of recombinant expression vector into Agrobacterium

[0115] Transform the correctly constructed recombinant expression vectors DBN100300 and DBN100300N (control sequence) into Agrobacterium GV3101 with the liquid nitrogen method, and the transformation conditions are: 100 μL of Agrobacterium GV3101, 3 μL of plasmid DNA (recombinant expression vector); placed in liquid nitrogen 10 minutes at 37°C in a warm water bath for 10 minutes; Inoculate the transformed Agrobacterium GV3101 in LB test tubes and culture at 28°C and 200rpm for 2 hours, and apply Rifampicin containing 50mg / L and 50mg / L of kanamycin on the LB plate until a positive single clone grows, pick a single clone and culture it and extract its plasmid, digest DBN100300 with restriction endonucleases PstI and SmaI, and carry out enzyme digestion verification. The restriction endonucle...

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Abstract

The present invention relates to a kind of herbicide resistance protein, its encoding gene and application, herbicide resistance protein comprises: (a) has the protein of the amino acid sequence composition shown in SEQ ID NO:2; Or (b) in (a) A protein derived from (a) in which the amino acid sequence in has been substituted and / or deleted and / or added with one or several amino acids and has herbicide resistance activity. The herbicide resistance protein of the present invention is particularly suitable for expression in plants and has wide resistance to herbicides, especially phenoxy auxin herbicides.

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): C12N9/02C12N15/53C12N15/84C12N5/10A01H5/00A01G7/06A01H6/46
CPCC12N9/0071C12N15/8209C12N15/8274C12N9/0069C12N15/8275C07K14/415C12N15/8277
Inventor 陶青吴业春牛晓广丁德荣
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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