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vip3a-1 gene of bacillus thuringiensis expression protein with high toxicity for asparagus caterpillar

A technology of Bacillus thuringiensis and beet armyworm, applied in the field of biochemistry, can solve problems such as poor control effect, threat to cotton production, and impact on cotton farmers' economic benefits

Inactive Publication Date: 2015-08-19
NORTHEAST AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, beet armyworms have been rampant, causing outbreaks and disasters. Many farmers frequently use pesticides but the control effect is not good, so they have to go to the fields to catch them every morning and evening.
For example, the rate of damaged plants in general cotton fields is about 60%, and the serious plots reach 100%. Once it happens, cotton farmers are often at a loss, which directly affects the economic benefits of cotton farmers and poses a serious threat to cotton production.

Method used

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  • vip3a-1 gene of bacillus thuringiensis expression protein with high toxicity for asparagus caterpillar
  • vip3a-1 gene of bacillus thuringiensis expression protein with high toxicity for asparagus caterpillar
  • vip3a-1 gene of bacillus thuringiensis expression protein with high toxicity for asparagus caterpillar

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Experimental program
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Effect test

Embodiment 1

[0025] 1. PCR amplification of genes:

[0026] The extracted Bt bacteria genomic DNA was amplified by PCR using the vip-5 / vip-3 primer pair, and the primer sequences are shown in Table 1. Referring to the N-terminal and C-terminal sequence homology of the coding region of vip3A gene published in GenBank, a pair of specific primers vip-5 / vip-3 were designed and synthesized, and NdeI and SalI restriction sites were introduced at the 5' end respectively (arrows indicated), for the amplification of the full-length gene.

[0027] Table 1: Primer sequences

[0028]

[0029] 2. PCR reaction system

[0030]

[0031]

[0032] 3. PCR reaction conditions

[0033] Table 2: PCR reaction conditions

[0034]

[0035] 4. DNA Recovery

[0036] (1) Use a sterilized scalpel to cut off the gel containing the target DNA fragment and place it in a 1.5mL centrifuge;

[0037] (2) Add sol buffer A with 3 times the volume of sol, and place it in a water bath at 50°C for about 10 minut...

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Abstract

The invention discloses a vip3A-1 gene of a bacillus thuringiensis expression protein with high toxicity for asparagus caterpillars, and belongs to the technical field of biochemistry. The expression protein is adopted for primary screening and secondary screening on bioactivity of asparagus caterpillars, the number of dead and live asparagus caterpillars after 7 days, the death rate and LC50 are calculated and corrected, the result shows that the medial lethal concentration of the vip3A-1 gene of the bacillus thuringiensis expression protein to asparagus caterpillars is 7.755mu g / g (5.683-9.979mu g / g) in secondary screening, and when the concentration of the expression protein is up to 81mu g / g, all asparagus caterpillars can be dead, so that the vip3A-1 gene of the bacillus thuringiensis expression protein has high toxicity for asparagus caterpillars, and is characterized in that the nucleotide sequence of the vip3A-1 gene of the bacillus thuringiensis expression protein with high toxicity for asparagus caterpillars is as shown in SEQ ID NO:1.

Description

technical field [0001] The invention relates to a vip3A-1 gene expressed by Bacillus thuringiensis which has high toxicity to beet armyworm and belongs to the technical field of biochemistry. Background technique [0002] Bacillus thuringiensis (Bt for short) belongs to the second category, Group 18 in the ninth edition of "Bergey's Bacteria Identification Manual", Gram-positive, and a species of Bacillus. During the formation of spores, one or more parasporal crystals with the same or different shapes are formed at one or both ends of the bacterium. The main component of this parasporal crystal is a protein with insecticidal activity, so it is also called insecticidal crystal protein (ICPs) or δ-endotoxin (δ-endotoxin). This insecticidal crystal protein is coded by cry gene and cyt gene. Now it has been found that most bacterial strains of Bacillus thuringiensis can produce multiple types of crystal protein, which is effective for Lepidoptera, Diptera, Coleoptera ( Coleop...

Claims

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Application Information

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IPC IPC(8): C12N15/32C07K14/325A01P7/04
Inventor 刘荣梅高继国李海涛张金波张杰束长龙赫福霞
Owner NORTHEAST AGRICULTURAL UNIVERSITY
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