Metabolic protein for weed-control agent, its gene, and its use

A technology of protein and herbicide, applied in the field of protein

Inactive Publication Date: 2007-01-10
SUMITOMO CHEM CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In addition, in the case of using herbicides, there are cases where it is difficult to distinguish cultivated plants from similar species of weeds to selectively control only weeds

Method used

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  • Metabolic protein for weed-control agent, its gene, and its use
  • Metabolic protein for weed-control agent, its gene, and its use
  • Metabolic protein for weed-control agent, its gene, and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0919] Example 1 Metabolism of Compound (II) by Microorganisms

[0920] (1) Metabolism of compound (II)

[0921] In ISP2 agar medium (1.0% (w / v) wort, 0.4% (w / v) yeast extract, 0.4% (w / v) glucose, 2.0% (w / v) agar, pH 7.3) Various microorganisms shown in Tables 1 and 2 were cultured. A loop amount of each microorganism was added to TGY medium (0.5% (w / v) tryptone, 0.5% (w / v) yeast extract, 0.1% (w / v) glucose, 0.01% (w / v) KH 2 PO 4 , pH 7.0) and incubated at 30°C with shaking for 2-4 days. One tenth milliliter (0.1 ml) of the obtained culture was added to 3 ml of sporulation broth (0.1% (w / v) meat extract, 0.2% (w / v) trypsin) containing 100 ppm of compound (II). % glucose, pH 7.1) for 7-8 days at 30°C with shaking. Fifty microliters (50 μl) of 2N HCl was added to the resulting culture and it was extracted with 3 ml of ethyl acetate. The ethyl acetate layer obtained was analyzed on HPLC. Decreasing the concentration of compound (II) (column retention time of 23.9 minutes)...

Embodiment 2

[0933] Example 2 Preparation of the protein (A1) of the present invention

[0934] (1) Preparation of crude cell extracts

[0935] A frozen stock of Streptomyces chromogenes IFO 12898 was added to 100 ml A medium (0.1% (w / v) glucose, 0.5% (w / v) tryptone, 0.5% (w / v) yeast) in a 500 ml Erlenmeyer flask Extract, 0.1% (w / v) dipotassium hydrogen phosphate, pH 7.0) and incubated at 30°C for 1 day with rotary shaking to obtain a pre-culture. Eight milliliters (8 ml) of the pre-culture was added to 200 ml A medium and incubated at 30°C for 2 days with rotary shaking in a 500 ml baffled shake flask. The cell pellet was recovered by centrifugation of the resulting culture (3,000 g, 5 min). The cell pellets were suspended in 100 ml B medium (1% (w / v) glucose, 0.1% beef extract, 0.2% (w / v) trypsin) containing 100 ppm compound (II) and incubated at 30°C. Incubate for 16 hours with reciprocating shaking in a 500 ml Sakaguchi flask. The cell pellet was recovered by centrifugation (3,000 ...

Embodiment 3

[0947] Example 3 Obtaining the DNA (A1) of the present invention

[0948] (1) Preparation of chromosomal DNA of Streptomyces dendrochromogenes IFO 12898

[0949] In 50 ml YEME medium (0.3% (w / v) yeast extract, 0.5% (w / v) peptone for bacteria, 0.3% (w / v) wort, 1.0% (w / v) glucose, 34% (w / v) w / v) sucrose and 0.2% (v / v) 2.5MgCl 2 ·6H 2 O) incubate Streptomyces chromogens IFO 12898 at 30°C for 1 to 3 days with shaking. Cells are recovered. The obtained cells were suspended in YEME medium containing 1.4% (w / v) glycine and 60 mM EDTA and further incubated with shaking for 1 day. Cells were recovered from the medium. After washing once with distilled water, it was resuspended in buffer (100 mM Tris-HCl (pH 8.0), 100 mM EDTA, 10 mM NaCl) at 1 ml per 200 mg of cells. Two hundred micrograms per milliliter (200 μg / ml) of egg white lysozyme was added. The cell suspension was incubated at 30°C for 1 hour with shaking. Additionally, 0.5% SDS and 1 mg / ml proteinase K were added. The ...

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Abstract

The invention provides DNAs encoding a weed controller metabolism protein selected from the following groups, which are useful in, for example, constructing a weed controller-tolerant plant. Proteins having an amino acid sequence represented by SEQ ID NO: 1, 2, 3, 108, 159, 160, 136, 137, 138, 215, 216, 217, 218, 219, 220, 221, 222, 223 or 224. Proteins having an amino acid sequence showing a sequence identity of 80% or more with one of the amino acid sequences represented by SEQ ID NOS: 1, 2, 3, 108, 159, 136, 137, 138, 215, 216, 217, 218, 219, 220, 221, 222, 223 or 224, or an amino acid sequence showing a sequence identity of 90% or more with one of the amino acid sequences represented by SEQ ID NO: 160, 215, 216, 218, 222 and 224, and being capable of converting the compound represented by the following formula II into the compound represented by the following formula III in the presence of an electron transfer system from an electron donor.

Description

technical field [0001] The present invention relates to a protein having the ability to metabolize herbicidal compounds (herbicide metabolizing protein), its gene and its application. Background technique [0002] The herbicides are used in the required amount of dilution when used. There are instances where excess remains. There are also situations in which the applied herbicide remains in the soil or plant residues shortly after its application. Initially, it was assumed that the safety of these herbicides had been checked, and that these small amounts of residual solutions or residues had little effect on the environment or the crops subsequently cultivated. However, if there is a method for converting the contained herbicidal compound into a compound of low herbicidal activity, a treatment to passivate the above-mentioned residual solution or residue, for example, can be carried out as required. [0003] In addition, in the case of using herbicides, there are cases in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C07K14/705C07K16/28C12N5/10A61K38/17A61K39/395G01N33/50G01N33/15C12P21/02C12P21/08A01H5/00C12R1/19C12R1/91A01HA61KC07KC12NC12PC12RG01N
Inventor 中岛宽树椋本藤夫高石昌直
Owner SUMITOMO CHEM CO LTD
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