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OBPs originated from Adelphocoris lineolatus and its coding gene

A kind of Lygus clover, gene technology, applied in the direction of genetic engineering, plant genetic improvement, chemicals for biological control, etc., can solve the problems of environmental damage, pollution, drug resistance of cotton bollworm, etc., to achieve improved efficiency, easy operation, low cost effect

Inactive Publication Date: 2012-04-04
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

Because Bt cotton is less effective against mirid gnats, the control of mirid gnats now mainly depends on chemical pesticides, which not only easily leads to the generation of insecticide resistance of Lepidoptera pests such as cotton bollworm, but also causes huge damage and pollution to the environment, so , it is necessary to develop cost-effective, environmentally friendly control methods

Method used

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  • OBPs originated from Adelphocoris lineolatus and its coding gene
  • OBPs originated from Adelphocoris lineolatus and its coding gene
  • OBPs originated from Adelphocoris lineolatus and its coding gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0050] Example 1: Extraction of antennal RNA, synthesis of first-strand cDNA by reverse transcription and synthesis of ds cDNA by LD-PCR

[0051] Get 200 pairs of antennae of Lygus alfalfa that just emerged, utilize the Trizol method to carry out the extraction of RNA, electrophoresis detects RNA quality (see figure 1 ), while the concentration of RNA measured by spectrophotometer was 618.8ng / μl. The first-strand cDNA and ds cDNA were synthesized by SMART technology and LD-PCR method. Finally, 1.5% agarose gel electrophoresis was used to detect the distribution of LD-PCR products. The principle of SMART is as follows figure 2 . The electrophoresis of LD-PCR products is shown in image 3 . The analysis results showed that the size of LD-PCR products ranged from 200bp to 2000bp, and the distribution range was relatively wide, indicating that the products included mRNAs of different genes, and there were several brighter bands, representing genes with relatively high expres...

Embodiment 2

[0052] Example 2: Digestion of ds cDNA by Sfi I and separation of ds cDNA by CHROMA SPIN-400 column

[0053] In order to connect the ds cDNA amplified by LD-PCR with the pDNR-LIB vector, the ds cDNA was digested with Sfi I endonuclease, and at the same time, the ds cDNA was separated by size using a CHROMA SPIN-400 column. In 16 sterile centrifuge tubes, one drop per tube. Finally, the size distribution of ds cDNA fragments detected by 1.5% agarose electrophoresis, such as Figure 4 , and finally according to the electrophoresis results, the ds cDNA and pDNR-LIB vector in tubes 7, 8, 9, and 10 were ligated.

Embodiment 3

[0054] Example 3: Electrotransformation of DH5α Escherichia coli cells with recombinant plasmids

[0055] In order to obtain higher conversion efficiency, electric conversion is used. The electroporation instrument is a product of Bio-Rad Company, and the electric shock parameters are set to 2000V, 200Ω. The electric shock time is 2-3 seconds.

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Abstract

The invention discloses odorant binding proteins (OBPs) originated from Adelphocoris lineolatus and its coding genes. According to the invention, an antenna cDNA library is constructed by utilizing the technology of SMART, and OBPs are effectively separated and identified from antenna of Adelphocoris lineolatus by using an information biological method. Since OBPs have a critical effect on olfactory recognition of insects, and the genes related in the invention can play an important role in biological control of insects.

Description

technical field [0001] The present invention relates to the technical field of using the olfactory system of insects to carry out pest biological control; further, the present invention relates to the method of using SMART technology to construct antennae cDNA library and combining bioinformatics methods to separate and identify insect odor binding protein OBPs and the isolated alfalfa blind Stinkbug odor-binding protein gene. Background technique [0002] Miridae, belonging to the Miridae family of Hemiptera, is an important class of pests in cotton production in my country. In 1997, my country began to commercially plant Bt cotton. By 2007, the planting area of ​​Bt cotton had reached 3.8 million hectares, accounting for 69% of the total cotton planting area in the country. The planting of Bt cotton has effectively controlled the harm of major lepidopteran pests such as cotton bollworm, and greatly reduced the use of chemical pesticides. Subsequently, a series of successi...

Claims

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Application Information

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IPC IPC(8): C07K14/435C12N15/12A01N61/00A01P19/00
Inventor 谷少华张永军张雪莹吴孔明郭予元
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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