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Farm crop fungus resistant polypeptides and method for making same

A technology for crops and fungi, which is applied in the field of anti-crop fungal polypeptides and its preparation, can solve the problems of increased drug resistance of crop fungi, large residues, unsatisfactory fungal curative effects, etc., and achieve good application prospects and specific targeting effects

Inactive Publication Date: 2013-06-26
畿晋庆堂国际生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The curative effect of antibacterial pesticides currently used on crops infected with fungi is not ideal; fungi are eukaryotic cells, and their similarity to human cells makes agricultural antifungal drugs such as tricyclazoles kill both fungal cells and human cells
The long-term use of chemical drugs such as cyclic azoles induces fungi to develop resistance to them, and farmers often use excessive doses of repeated drugs to suppress fungal infections of crops, which further aggravates the resistance of fungi in crops; The amount of residues in the environment and agricultural products is increasing, so humans have been working on the development of highly efficient and pollution-free antifungal pesticides

Method used

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  • Farm crop fungus resistant polypeptides and method for making same
  • Farm crop fungus resistant polypeptides and method for making same
  • Farm crop fungus resistant polypeptides and method for making same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Construction of a plasmid expressing an anti-crop fungal polypeptide and preparation of a recombinant anti-crop fungal polypeptide

[0020] The original plasmid was the pET-15b commercial plasmid loaded with colicin Ia and Immunity protein genes (the size of the plasmid was 6.3 kb, purchased from Novagen, and the above genes were loaded by our laboratory), and the double-stranded oligonucleotide point mutation technique ( QuickChange TM Kit, Strategene Company) inserted the gene encoding Candida albicans pheromone (the nucleotide sequence described in SEQ ID NO.2 in the sequence listing) into the I626 site of the colicin Ia gene, and prepared the anti-crop fungal polypeptide A recombinant plasmid pCHCCACH1 (as shown in Figure 1). The recombinant plasmid is transfected into the E.coli BL21 (DE3) plyS engineering bacteria to prepare the polypeptide, and the anti-crop fungal polypeptide described in SEQ ID NO.5 in the sequence table is obtained, which is called...

Embodiment 2

[0051] Example 2 Construction of a plasmid expressing an anti-crop fungal polypeptide and preparation of a recombinant anti-crop fungal polypeptide

[0052] The original plasmid is the pET-15b commercial plasmid loaded with colicin Ia and Immunity protein genes (plasmid size 6.3kb, purchased from Novagen, the above-mentioned genes are loaded by our laboratory), through double-stranded oligonucleotide point mutation technology ( QuickChange TM Kit, Strategene Company) inserted the gene encoding Candida albicans pheromone (nucleotide sequence described in SEQ ID NO.2 in the sequence listing) before the S1 site of colicin Ia, and prepared the anti-crop fungal polypeptide Yet another recombinant plasmid pCHCCACH2 (as shown in FIG. 2 ). The recombinant plasmid was transfected into the E.coli BL21 (DE3) plyS engineering bacteria to prepare the polypeptide, and the anti-crop fungal polypeptide described in SEQ ID NO.7 in the sequence table was obtained, which was called "anti-crop ...

Embodiment 3

[0070] Example 3 Anti-crop fungus polypeptide to the in vitro inhibition test of crop fungi

[0071] Test 1: The fungus was the fungus Pyricularia oryzae isolated from the field by the Plant Protection Station of the Yucheng District Agriculture Bureau, Ya'an City, Sichuan Province. Take 5 microliters of the bacterial solution (10 8 CFU / ml level of bacteria) evenly spread on the surface of Sapporo's solid medium (Hangzhou Microbial Reagent Factory), and then place a small plastic cup to absorb 50 μl of each treatment solution on the surface of the medium, each treatment solution is the concentration of tricyclazole 5 mg / ml, tricyclazole concentration 50 μg / ml, anti-crop fungal polypeptide 2 concentration 5 μg / ml, anti-crop fungal polypeptide 1 concentration 5 μg / ml. The medium was cultured at 35°C for two days, and the results are shown in Figure 3. The results showed that only the treatment solution with a concentration of 5 mg / ml of tricyclazole and the treatment solution w...

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Abstract

The present invention discloses a novel anti-crop-epiphyte polypeptide and a preparation method thereof. A recombined anti-crop-epiphyte polypeptide is formed by linking gene which codes white candida pheromone and the gene which codes colicin. Compared with the prior anti-crop-epiphyte medicine, the novel anti-crop-epiphyte polypeptide of the present invention has the advantages that the novel anti-crop-epiphyte polypeptide directly forms ionic passage at epiphytic cell membrane to kill the epiphyte, so the killing efficiency is thousands times stronger than the prior anti-crop-epiphyte medicine; the epiphyte can hardly amend the geometrical damage at the cell membrane caused by the polypeptide by the mutation-expression form, so the polypeptide can not induce the epiphyte producing the traditional drug resistance; the polypeptide is a biological preparation and has target killing towards the epiphyte, so the novel anti-crop-epiphyte polypeptide does not have the toxic and side effects of the prior anti-crop-epiphyte medicine.

Description

technical field [0001] The present invention relates to a recombinant gene, recombinant plasmid, polypeptide and preparation method of anti-crop fungus polypeptide. Background technique [0002] Fungal infection is a common hazard to crops. For example, the fungus Pyrrhiza oryzae that causes rice blast causes huge economic losses to my country's rice production every year, and is extremely harmful. With the acceleration of urbanization in the vast agricultural areas of our country, the pollution and deterioration of the ecological environment are increasing day by day, and the incidence of crop diseases has risen sharply. The ecological prevention and control of crop diseases has become a major problem that we must face and solve. [0003] The curative effect of antibacterial pesticides currently used on crops infected with fungi is not ideal; fungi are eukaryotic cells, and their similarity to human cells makes agricultural antifungal drugs such as tricyclazoles kill human ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/31C12N15/63C07K19/00A01P3/00
Inventor 丘小庆
Owner 畿晋庆堂国际生物技术有限公司
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