Migratory locust protein tyrosine kinase PTK and coding gene and application thereof

A tyrosine kinase and coding gene technology is applied in the field of migratory locust protein tyrosine kinase PTK and its coding gene and application to achieve the effect of understanding the diapause mechanism of migratory locusts

Inactive Publication Date: 2018-05-22
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

Studies on mammals have shown that protein tyrosine kinase (PTK) can participate in the balance regulation of intracellular protein tyrosine, but whether PTK is involved in the regulation of diapause in insects has not been reported

Method used

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  • Migratory locust protein tyrosine kinase PTK and coding gene and application thereof
  • Migratory locust protein tyrosine kinase PTK and coding gene and application thereof
  • Migratory locust protein tyrosine kinase PTK and coding gene and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0069] Embodiment 1, acquisition of migratory locust protein tyrosine kinase PTK and its coding gene

[0070] 1. Extraction of total RNA from migratory locusts

[0071] use RNA isolation reagents were used to extract RNA from migratory locust tissue samples. Specific steps are as follows:

[0072] 1) Put a 2ml homogenizer in an oven at 160°C for 3 hours, and cool to room temperature for later use.

[0073] 2) Put the homogenizer on ice, add 1ml RNA isolation reagent and 100-200mg migratory locust tissue, ground.

[0074] 3) Transfer the homogenate to a 1.5ml centrifuge tube and let stand at room temperature for 5 minutes. Centrifuge at 13000r for 5min at 4°C.

[0075] 4) Transfer the supernatant to a clean 1.5ml centrifuge tube, add 200μl chloroform, and vortex for 15s. Place at room temperature for 5 minutes. Centrifuge at 13000r for 10min at 4°C.

[0076] 5) Transfer 400 μl of supernatant to a new 1.5 ml centrifuge tube, add 200 μl of chloroform, and vortex for 30...

Embodiment 2

[0109] Embodiment 2, the dsRNA of migratory locust protein tyrosine kinase gene PTK and its application in controlling migratory locust

[0110] 1. Synthesis of dsRNA

[0111] Using T7RiboMAX TM The Express RNAi System Kit synthesizes dsRNA. Specific steps are as follows:

[0112] 1) Synthesis of dsRNA primers

[0113] Design primers based on the cloned gene fragments, amplify the target fragment to about 600bp, and introduce T7 promoter at the 5' end of the primers. The primer sequences are as follows:

[0114]PTK-2F: 5'-TAATACGACTCACTATAGGACGATTCACAGGAACATTGGTG-3';

[0115] PTK-2R: 5'-TAATACGACTCACTATAGGCATCTTGACAGCGACATCTATGG-3'.

[0116] 2) Preparation of DNA template

[0117] The bacterial liquid plasmid was extracted with a kit, and the plasmid containing the gene fragment (recombinant vector in Example 1) was used as a template, and PTK-2F and PTK-2R were used for PCR amplification to obtain the target fragment containing the T7 promoter sequence.

[0118] The ...

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Abstract

The invention discloses a migratory locust protein tyrosine kinase PTK and coding gene and application thereof. The method comprises the following steps: at first, cloning migratory locust protein tyrosine kinase gene PTK from the migratory locust, and designing a primer for the migratory locust protein tyrosine kinase gene PTK; synthesizing dsRNA for interfering migratory locust protein tyrosinekinase gene PTK, and then importing the dsRNA into the migratory locust to perform RNAi on the migratory locust protein tyrosine kinase gene PTK by using an injection method. The result shows that themigratory locust protein tyrosine kinase gene PTK can regulate and control the diapause of the migratory locust. The invention provides the theoretical basis for deeply understanding migratory locustdiapause mechanism and creating the new biological pesticide preparation.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to migratory locust protein tyrosine kinase PTK and its coding gene and application. Background technique [0002] Migratory locust is a major pest in agriculture and animal husbandry. It survives the winter with embryo diapause, and belongs to facultative diapause insect. Diapause-inducing external conditions include temperature, photoperiod and so on. The mechanisms of diapause regulation that have been reported so far include molecular regulation, hormone regulation, circadian clock regulation and energy regulation. More recognized is the regulation of insulin signaling. This signaling pathway is associated with multiple features of diapause, such as suppressed reproduction, prolonged lifespan, suppressed metabolism, fat accumulation, and enhanced stress tolerance. Insulin signaling can be regulated by tyrosine phosphorylation of signaling molecules in the insulin sign...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/54A01N57/16A01P7/04
CPCA01N57/16C12N9/1205C12Y207/01112
Inventor 涂雄兵张泽华崔栋楠赵海龙王广君农向群
Owner INST OF PLANT PROTECTION CHINESE ACAD OF AGRI SCI
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