Locust uridine diphosphate glucuronic acid transferase gene and application thereof

A technology of uridine diphosphate glucose and transferase, applied in the direction of transferase, application, genetic engineering, etc., can solve the problems of unfavorable agricultural environment green and sustainable development, and achieve the effect of reducing selectivity and fitness

Active Publication Date: 2019-06-21
LINYI UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, traditional chemical control methods are mostly used for the prevention and control of Asian

Method used

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  • Locust uridine diphosphate glucuronic acid transferase gene and application thereof
  • Locust uridine diphosphate glucuronic acid transferase gene and application thereof
  • Locust uridine diphosphate glucuronic acid transferase gene and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 Obtaining of UGT encoding gene of uridine diphosphate glucuronyl transferase from locust asiatica

[0022] 1. Extraction of total RNA from locust Asiatica

[0023] use The separation reagent extracts the total RNA from the sample of A. asiatica. Specific steps are as follows:

[0024] 1) Put the homogenizer on ice and add 1mL Separate the reagents and 100-200mg of Asiatic locust tissue and grind.

[0025] 2) Transfer the homogenate to a 1.5mL centrifuge tube, let it stand at room temperature for 5 minutes, and centrifuge at 13000r for 5 minutes at 4°C.

[0026] 3) Transfer the supernatant to a clean 1.5 mL centrifuge tube, add 200 μL of chloroform, and vortex for 15 seconds. Place it at room temperature for 5 minutes, and centrifuge at 13000r for 10 minutes at 4°C.

[0027] 4) Aspirate 400 μL of supernatant and transfer to a new 1.5 mL centrifuge tube, add 200 μL of chloroform, vortex for 30 seconds, place at room temperature for 5 minutes, and centrifuge at 13000r f...

Embodiment 2

[0060] Example 2 Synthesis of dsRNA and its application in pest control

[0061] 1. Synthesis of dsRNA

[0062] Adopt T7 RiboMAX TM Express RNAi System kit synthesizes dsRNA. Specific steps are as follows:

[0063] 1) Synthesis of dsRNA primers

[0064] Design primers based on the cloned gene fragments, and introduce T7 promoter at the 5'end of the primers. The primer sequence is as follows:

[0065] UGT-2F: 5'-GGATCCTAATACGACTCACTATAGGGCTGTTCTTCGCTCTAAACC-3' (SEQ ID NO: 3);

[0066] UGT-2R: 5'-GGATCCTAATACGACTCACTATAGGGGCAAATGTATTCCACCAAC-3' (SEQ ID NO: 4).

[0067] 2) Preparation of DNA template

[0068] The bacterial liquid plasmid of the positive cloned strain in Example 1 was extracted with the kit, and the plasmid was used as a template to perform PCR amplification using UGT-2F and UGT-2R to obtain a target fragment containing the T7 promoter sequence, with a size of 520 bp.

[0069] The PCR reaction system is as follows: plasmid 1μL, dNTP 4μL, 10×Buffer 5μL, UGT-2F 1μL, UGT-2R 1...

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PUM

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Abstract

The invention discloses a locust uridine diphosphate glucuronic acid transferase gene. The nucleotide sequence of the gene is as shown in SEQ ID NO: 1. According to the gene, a primer is designed to synthesize dsRNA for interfering the uridine diphosphate glucuronic acid transferase gene, the dsRNA is introduced into the uridine diphosphate glucuronic acid transferase gene to perform RNA (ribonucleic acid) interference, food selectivity and conformity of locusts can be remarkably reduced, a novel method is provided for green and sustainable prevention and control of injurious insects, and technical support is provided for innovation of novel biological pesticides.

Description

Technical field [0001] The invention relates to the field of biotechnology, and more specifically to a locust uridine diphosphate glucuronyl transferase gene and its application. Background technique [0002] Pest damage to crops will reduce productivity and endanger food security. Traditional pest control methods are mostly based on chemical control, but over-reliance on the use of chemical pesticides can easily cause pest resistance, resurgence, and serious residual poisons, which will seriously damage the ecological environment and the quality and safety of agricultural products. Therefore, it is particularly important to develop and create green, efficient and sustainable pest control technologies. [0003] Oedaleus asiaticus Bey-Bienko is one of the most serious dominant species of locusts in northern my country. It can feed on gramineous grasses and food crops. It likes to feed on Stipa kleinii, which seriously damages the agricultural production in the north. Biological dis...

Claims

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Application Information

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IPC IPC(8): C12N15/54C12N9/10C12N15/89A01K67/033A01N37/46A01P7/04
CPCY02A50/30
Inventor 黄训兵王月月刘文
Owner LINYI UNIVERSITY
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