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Bacillus thuringiensis insecticidal crystal protein gene and application

A technology of crystal protein and thuringiensis spore, which is applied in application, insecticide, genetic engineering, etc., to achieve the effect of unique insecticidal properties, low production and use costs, and safety for humans, animals and plants

Inactive Publication Date: 2008-02-20
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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Problems solved by technology

[0005] However, most of the insecticidal crystal proteins encoded by these genes kill pests such as Lepidoptera or Coleoptera. So far, there is no relevant Bacillus thuringiensis (Bt) insecticidal crystal protein gene and its coding for killing locusts in the world. Reports of insecticidal crystal proteins

Method used

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  • Bacillus thuringiensis insecticidal crystal protein gene and application

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Embodiment Construction

[0021] 1. Extraction of the total DNA of the locust-resistant Bacillus thuringiensis strain:

[0022] The activated BTH-13 strain was inoculated on the plate and cultured at 30°C for 8h. The bacterial cells were washed with 20 mmol / l Tris.cl (pH8.0) buffer solution, centrifuged at 5000 rpm for 5 min, and the bacterial cells were collected. Add 1 volume of TES solution (containing 15 mg / ml lysozyme), incubate at 37°C for 2 h, add 2 volumes of solution II, mix well, invert slightly, add proteinase K to a final concentration of 70 ug / ml. Incubate at 37°C for 2h until the solution becomes clear. Add 1.5 times the volume of Solution III, mix well, centrifuge at 12000rpm for 15min, and collect the supernatant. Add an equal volume of phenol / chloroform (1:1), mix thoroughly, and invert slightly for at least 5 minutes to remove protein. Centrifuge at 12,000 rpm at room temperature for 5 minutes, collect the supernatant, add an equal volume of chloroform / isoamyl alcohol (24:1), and r...

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Abstract

The invention discloses an insecticidal crystal protein gene of bacillus thuringiensis and the application thereof; the insecticidal protein coded by a DNA sequence of the insecticidal crystal protein gene shown in SEQ ID No.1 is an amino acid sequence shown in SEQ ID No.2. The insecticidal protein coded by the gene has specific insecticidal function to orthopterous oriental migratory locust, the locust insecticide prepared by the gene has good prevention effects used for preventing and curing those locusts harmful to graminaceous crops such as grain, wheat, corn, broomcorn and pasture etc., and is safe to human and animal and plant, has low production and using cost, does not cause environmental pollution, thereby having great application value.

Description

technical field [0001] The invention relates to a locust-resistant Bacillus thuringiensis insecticidal crystal protein gene. The insecticidal protein encoded by the gene is active and toxic to Orthoptera migratory locusts, etc. The locusticide prepared by engineering bacteria constructed based on the gene can be used for The control of orthoptera pests such as locusts and the use of the gene in insect-resistant transgenic plants. Background technique [0002] Locusts are major agricultural pests worldwide. Locust plagues, floods, and droughts often alternate or accompany each other and become the three major natural disasters in human history. Since the 1980s, global locust disasters have entered a stage of frequent occurrence. Except for Antarctica, locust disasters have occurred on all continents. The locust plague in my country has also increased. Since 1997, locust plagues have broken out in a large area for several consecutive years in my country, and the highest popu...

Claims

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Application Information

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IPC IPC(8): C12N15/32C07K14/325A01N63/00A01P7/04
Inventor 高梅影宋玲莉戴顺英
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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