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Protein tyrosine phosphatase PTP1B of Locusta migratoria and encoding gene and application thereof

A technology of tyrosine phosphatase and coding gene, applied in locust protein tyrosine phosphatase PTP1B and its coding gene and application field

Inactive Publication Date: 2018-08-07
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 also shown that protein tyrosine phosphatase 1B (Proteintyrosinephosphatase 1B, PTP1B) can participate in the balance regulation of intracellular protein tyrosine, but whether PTP1B is involved in the regulation of diapause in insects has not been reported

Method used

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  • Protein tyrosine phosphatase PTP1B of Locusta migratoria and encoding gene and application thereof
  • Protein tyrosine phosphatase PTP1B of Locusta migratoria and encoding gene and application thereof
  • Protein tyrosine phosphatase PTP1B of Locusta migratoria and encoding gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Example 1, the acquisition of migratory locust protein tyrosine phosphatase PTP1B and its coding gene

[0069] 1. Extraction of total RNA from migratory locusts

[0070] use RNA isolation reagents were used to extract RNA from migratory locust tissue samples. Specific steps are as follows:

[0071] 1) Put a 2ml homogenizer in an oven at 160°C for 3 hours, and cool to room temperature for later use.

[0072] 2) Put the homogenizer on ice, add 1ml RNA isolation reagent and 100-200mg migratory locust tissue, ground.

[0073] 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.

[0074] 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.

[0075] 5) Transfer 400 μl of supernatant to a new 1.5 ml centrifuge tube, add 200 μl of chloroform, and vorte...

Embodiment 2

[0108] Example 2, the dsRNA of migratory locust protein tyrosine phosphatase gene PTP1B and its application in controlling migratory locust

[0109] 1. Synthesis of dsRNA

[0110] Using T7RiboMAX TM The Express RNAi System Kit synthesizes dsRNA. Specific steps are as follows:

[0111] 1) Synthesis of dsRNA primers

[0112] 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:

[0113] PTP1B-2F: 5'-TAATACGACTCACTATAGCCTTCTGCCTTGTAGATTCTTGC-3';

[0114] PTP1B-2F: 5'-TAATACGACTCACTATAGGCTTCCTCCTCTTCTTGCTGGAC-3'.

[0115] 2) Preparation of DNA template

[0116] 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 to perform PCR amplification using PTP1B-2F and PTP1B-2R to obtain the target fragment containing the T7 promoter sequence.

[011...

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PUM

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Abstract

The invention discloses protein tyrosine phosphatase PTP1B of Locusta migratoria and an encoding gene and application thereof. DsRNA of protein tyrosine phosphatase gene PTP1B of protein tyrosine phosphatase is cloned from protein tyrosine phosphatase; primers are designed for the protein tyrosine phosphatase gene PTP1B of Locusta migratoria; dsRNA for interfering the protein tyrosine phosphatasegene PTP1B of Locusta migratoria is synthesized; the dsRNA can be imported into the protein tyrosine phosphatase gene PTP1B of Locusta migratoria to carry out RNAi (RNA interference). The results showthat the protein tyrosine phosphatase PTP1B of Locusta migratoria can regulate the diapause of Locusta migratoria. Theoretical basis is provided for the deeper understanding of the diapause mechanismof Locusta migratory and the development of biopesticide preparations.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to migratory locust protein tyrosine phosphatase PTP1B 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 insuli...

Claims

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Application Information

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IPC IPC(8): C12N9/16C12N15/55C12N15/85A01K67/033
CPCA01K67/0339A01K2217/058A01K2227/706C12N9/16C12Y301/03048
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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