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Insecticidal gene cry2Ah-like with high toxicity on lepidoptera pest and application thereof

A kind of insecticidal gene and Lepidoptera technology, applied in the fields of application, insecticide, genetic engineering, etc., can solve the problems of single and increased pest resistance, achieve high virulence, delay the occurrence of insecticide resistance, and enrich insecticidal gene resources Effect

Active Publication Date: 2013-07-17
INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Since the current commercialized transgenic insect-resistant crops have relatively single types of insect-resistant genes, such a large-scale promotion of planting has the risk of reducing pest refuges and increasing pest resistance.

Method used

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  • Insecticidal gene cry2Ah-like with high toxicity on lepidoptera pest and application thereof
  • Insecticidal gene cry2Ah-like with high toxicity on lepidoptera pest and application thereof
  • Insecticidal gene cry2Ah-like with high toxicity on lepidoptera pest and application thereof

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Embodiment Construction

[0022] The present invention will be further described in detail below in conjunction with the examples.

[0023] Bt plasmid extraction

[0024] RNaseA (10mg / ml): Weigh 0.1g of RNaseA and dissolve it in 10ml of 10mM Tris·HCl (pH7.5), 15mM NaCl, boil at 100°C for 15min, cool in an ice bath, and store at -20°C for later use.

[0025] The SC6H8 strain is preserved in the applicant's laboratory and can be released to the public.

[0026] 1. Incubate 5 ml of SC6H8 bacterial broth overnight.

[0027] 2. Collect the cells by centrifugation, suspend the cells with a 25% sucrose solution containing 30 mg / ml lysozyme, the total volume is 200 μl, and incubate at 37° C. for 15 mins.

[0028] 3. Add 400 μl of alkaline lysis solution (3% SDS, 0.2N NaOH), mix slowly immediately, and incubate at room temperature for 5 mins.

[0029] 4. Add 300 μl ice-cold 3M sodium acetate (pH 4.8), mix immediately, and centrifuge at 12000g for 10mins.

[0030] 5. Take the supernatant, add an equal volume...

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Abstract

The invention relates to an insecticidal gene cry2Ah-like with high toxicity on lepidoptera pests and an application thereof, and belongs to the technical field of biology. The protein sequences of insecticidal proteins Cry2Ah-like, Cry2Ah-likesp and Cry2Ah-likevp with high toxicity on the lepidoptera pests are respectively shown as SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3. The new gene provided by the invention achieves the high toxicity on cotton bollworm and corn borer, enriches the insecticidal gene resources, shows the toxicity on relative pests by being applied to transform microbes and overcomes and postpones the drug resistance of the pests on engineering bacteria.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to new insecticidal genes cry2Ah-like, cry2Ah-likesp, cry2Ah-likevp with high virulence of Bacillus thuringiensis to lepidopteran pests and applications thereof. Background technique [0002] Insect pests are one of the important causes of crop yield reduction, and reducing the loss of pests is an important way to increase the yield of food and feed crops. According to statistics, the total production of food and feed crops in the world loses 14% of the total annual output due to insect pests, and the economic losses directly caused to agricultural production are as high as hundreds of billions of dollars. The annual losses caused by insect pests in my country reduce rice production by 10%, wheat production by 20%, and cotton production by more than 30%. The use of chemical pesticides and biological pesticides and other control methods can certainly reduce the damage of pests to crops,...

Claims

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Application Information

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IPC IPC(8): C07K14/325C12N15/32C12N15/63A01N47/44A01P7/04C12N15/70C12N1/21A01H5/00C12R1/07
Inventor 束长龙张杰耿丽丽宋福平彭琦黄大昉
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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