Fusion gene of compound antibody type with glyphosate and glufosinate, encoded protein and application thereof

A glyphosate-resistant protein and fusion gene technology, which is applied in the fields of application, fusion polypeptide, genetic engineering, etc., can solve extensive and other problems

Inactive Publication Date: 2017-01-11
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Human beings have been controlling weeds since the beginning of agricultural produ

Method used

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  • Fusion gene of compound antibody type with glyphosate and glufosinate, encoded protein and application thereof
  • Fusion gene of compound antibody type with glyphosate and glufosinate, encoded protein and application thereof
  • Fusion gene of compound antibody type with glyphosate and glufosinate, encoded protein and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 Construction of glyphosate and glufosinate-ammonium composite resistance fusion protein expression vector

[0028]The fusion gene formed by the fusion of the glufosinate-resistant gene bar and the glyphosate-resistant gene GAT is named bar-GAT gene, and the encoded protein polypeptide sequence is shown in SEQ ID NO:3. The fusion gene formed by the fusion of glyphosate-resistant gene CP4 and glufosinate-resistant gene pat is named CP4-pat gene, and the encoded protein polypeptide sequence is shown in SEQ ID NO:4.

[0029] bar-GAT gene (nucleotide sequence as shown in the 11th-1021st base in SEQ ID NO: 1), artificially synthesized with a BamHI restriction site at the 5' end, and artificially synthesized at the 3' end terminator, and a KpnI restriction site is set, and the nucleotide sequence is shown in SEQ ID NO:1. CP4-pat gene (nucleotide sequence as shown in the 224-2164th base in SEQ ID NO: 2) and the chloroplast transit peptide CTP at the 5' end of the ...

Embodiment 2

[0038] Embodiment 2, transformation of rice

[0039] The method of obtaining transgenic rice is to adopt the existing technology (Lu Xiongbin, Gong Zuxun (1998) Life Science 10: 125-131; Liu Fan et al. (2003) Molecular Plant Breeding 1: 108-115). The mature and plump seeds of "Xiushui-134" were dehulled, and callus was induced as transformation materials. The Agrobacterium plates of the vectors pCambia1300-pUBI-bar-GAT-p35S-1174 and pCambia1300-pUBI-CP4-PAT-p35S-1174 constructed in Example 1 were respectively taken. Pick a single colony to inoculate and prepare Agrobacterium for transformation. Put the callus to be transformed into the Agrobacterium bacterium solution with an OD of about 0.6 (preparation of the Agrobacterium bacterium solution: inoculate the Agrobacterium into the medium, and cultivate it at 28°C until the OD is about 0.6; the composition of the medium: 3g / L K 2 HPO 4 , 1g / LNaH 2 PO 4 , 1g / LNH 4 Cl, 0.3g / L MgSO 4 ·7H 2 O, 0.15g / L KCl, 0.01g / L CaCl 2...

Embodiment 3

[0040] Example 3, fusion protein transgenic rice can resist glufosinate-ammonium

[0041] The T0 generation plants of the transgenic rice plants prepared by the method in Example 2 were transplanted into the greenhouse, and the herbicide resistance of the transgenic rice plants and the non-transgenic recipient control plant "Xiu Shui-134" were compared and analyzed. We obtained 128 transgenic lines (named BG) and 155 transgenic lines transgenic pCambia1300-pUBI-CP4-pat–p35S-1174 vector (named Carry out different concentrations of glufosinate-ammonium resistance determination for GC), and the resistance effect is as shown in table 1:

[0042] Table 1*:

[0043]

[0044] *Note: Table 1 shows the resistance test of rice plants sprayed with different concentrations of glufosinate-ammonium 15 days after sowing. 10 days after spraying glufosinate-ammonium, the plants with no significant difference in plant height, number of leaves and growth vigor compared with the plants spray...

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Abstract

The present invention discloses a fusion gene of compound antibody type with glyphosate and glufosinate, encoded protein and application thereof, The fusion gene composes of a glyphosate-tolerant encoded-protein gene and a glufosinate-tolerant encoded-protein gene. The glyphosate encoded gene is one of the following: CP4, aroA, G7, G10, GOX or GAT; the glufosinate encoded gene is bar or pat. The compound protein can give compound antibodies to the plants, glyphosate and glufosinate; it can be used to produce transgene plants with the antibody property of glyphosate and glufosinate. The provided compound protein with antibody properties of glyphosate and glufosinate can be used in herbicides for monocotyledonous plants and dicotyleons; it is mainly used in herbicides for insect resistant corn, rice, soybean, wheat and rape.

Description

[0001] (1) Technical field [0002] The invention relates to a fusion gene with combined resistance to glyphosate and glufosinate-ammonium, an encoded protein, and an application of the fusion protein. [0003] (2) Background technology [0004] Weeds and pests are the most important biological factors affecting yield in agricultural production. How to reduce the control cost of weeds and pests, improve the control efficiency, and reduce the environmental pollution during the control process are the most critical issues in agricultural science research. [0005] Weeds are produced along with human production activities, and their existence is the result of long-term adaptation to climate, soil, crops, farming systems and social factors and competition with cultivated crops. Human beings have been controlling weeds since the beginning of agricultural production, but weed control at that time was only an extremely extensive primary labor. With the development of science, especi...

Claims

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Application Information

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IPC IPC(8): C12N15/62C07K19/00C12N15/54C12N15/82A01H5/00
CPCC12N9/1025C07K2319/00C12N9/1029C12N9/1092C12N15/8275C12N15/8277C12Y203/01183C12Y205/01019
Inventor 张先文王东芳沈志成
Owner ZHEJIANG UNIV
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