Herbicide resistance protein, coding gene and application thereof

A resistance gene and herbicide technology, applied in herbicides and algicides, genetic engineering, plant genetic improvement, etc., can solve the problem of not finding the expression level of 24DT07 herbicide resistance protein, herbicide tolerance report, etc. Achieve the effect of improving flexibility and high flexibility

Active Publication Date: 2014-12-17
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 24DT07 herbicide resistance protein in plants and its tolerance to herbicides

Method used

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  • Herbicide resistance protein, coding gene and application thereof
  • Herbicide resistance protein, coding gene and application thereof
  • Herbicide resistance protein, coding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

no. 1 example 、24D

[0098] The first embodiment, the acquisition and synthesis of 24DT07 gene sequence

[0099] 1. Obtain the 24DT07 gene sequence

[0100] The amino acid sequence (293 amino acids) of the 24DT07 herbicide resistance protein is shown in SEQ ID NO: 2 in the sequence listing; the amino acid sequence (293 amino acids) corresponding to the 24DT07 herbicide resistance protein is obtained according to the plant-preferred codons. amino acids) nucleotide sequence (882 nucleotides), as shown in SEQ ID NO:1 in the sequence listing.

[0101] 2. Synthesize the above-mentioned 24DT07 nucleotide sequence

[0102] The 24DT07 nucleotide sequence (shown as SEQ ID NO: 1 in the sequence listing) was synthesized by Nanjing GenScript Biotechnology Co., Ltd.; the 5' of the synthesized 24DT07 nucleotide sequence (SEQ ID NO: 1) A SpeI restriction site is also connected to the end, and a KasI restriction site is also connected to the 3' end of the 24DT07 nucleotide sequence (SEQ ID NO: 1).

[0103] At ...

no. 2 example

[0106] The second embodiment, construction of Arabidopsis recombinant expression vector and transformation of recombinant expression vector into Agrobacterium

[0107] 1. Construction of Arabidopsis thaliana recombinant cloning vector DBN01-T containing 24DT07 nucleotide sequence

[0108] The synthetic 24DT07 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 initiation codon; SP6 is the SP6 RNA polymerase promoter; T7 is the T7 RNA polymerase promoter; 24DT07 is 24DT07 Nucleotide sequence (SEQ ID NO: 1); MCS is multiple cloning site).

[0109] Then, the recombinant cloning vector DBN01-T was tra...

no. 3 example

[0125] The third embodiment, the acquisition of Arabidopsis plants transferred to 24DT07 nucleotide sequence

[0126] 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. Seeds were germinated and plants were grown in a greenhouse under long-day conditions (16 h light / 8 h dark) at constant temperature (22 °C) constant humidity (40-50%) and light intensity of 120-150 μmol / m2 sec. Start by watering the plants with Hoagland's nutrient solution, followed by deionized water, keeping the soil moist but not soggy.

[0127] Transform Arabidopsis using the flower soak method. Inoculate one or more 15-30ml precultures of YEP broth ...

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Abstract

Some embodiments of the present invention can include herbicide-resistant proteins, coding genes, and uses thereof. Certain herbicide-resistant proteins can comprises: (a) a protein consisting of an amino acid sequence shown in SEQ ID NO: 2; or (b) a protein consisting of an amino acid sequence at least 90% identical to that set forth in SEQ ID NO: 2. Other herbicide-resistant proteins of this disclosure can be suitable for expression in plants, can have resistance to herbicides (e.g., to phenoxy auxinherbicides), or both.

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): C12N9/02C12N15/53C12N15/63C12N5/10C12N1/13C12N1/19C12N1/21C12N7/01A01K67/033A01H5/00A01N47/44A01P13/00
CPCC07K14/001C07K14/415
Inventor 吴业春陶青丁德荣王登元刘海利宋肖倩邢甜甜杜立国尚松松鲍晓明
Owner BEIJING DABEINONG BIOTECHNOLOGY CO LTD
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