Chitinase ChiCl for deactivating varroa destructor and application thereof

A technology for killing Varroa dissimilar and chitinase, which is applied in the fields of application, acaricide, enzyme, etc.

Inactive Publication Date: 2012-05-23
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, no reports have been found on the use of chitinase to control Varroa dissimilarus in honeybees

Method used

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  • Chitinase ChiCl for deactivating varroa destructor and application thereof
  • Chitinase ChiCl for deactivating varroa destructor and application thereof
  • Chitinase ChiCl for deactivating varroa destructor and application thereof

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Embodiment 1

[0024] The Serratia bacterium GEI Serratia marcescens GEI described in the present invention is obtained by the inventor from the midgut of bee worker bees in Conghua. This bacterium was preserved in the China Center for Type Culture Collection (CCTCC, address : Wuhan University, Wuhan, China), the deposit number is CCTCC M 209097.

[0025] 1. Cloning and sequencing of the coding gene of chitinase ChiC1.

[0026] 1. Culture of Serratia GEI Serratia marcescens GEI

[0027] Pick a single colony of Serratia GEI Serratia marcescens GEI from the LB plate and inoculate it into 4mL LB liquid medium, and cultivate it overnight with regular shaking (28°C, 200r / min); Culture medium in a 100mL Erlenmeyer flask, cultured to OD620≈2.

[0028] 2. Extraction of Bacterial Genomic DNA

[0029] Genomic DNA of Serratia strain GEI was extracted using a conventional kit (TIANGEN Bacterial Genome Extraction Kit, Beijing TIANGEN Company).

[0030] 3. Primer Design

[0031] According to the chit...

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Abstract

The invention discloses a chitinase ChiCl for deactivating a varroa destructor and application thereof. In the invention, The chitinase ChiCl for deactivating the varroa destructor is prepared by cloning a gene base sequence from a genome DNA of serratia marcescens GEI, and applying a procaryon expression carrier pET-32a(+) to express the gene in colon bacillus. Purification, renaturation and bioassay are carried out on a protein coded by the gene, a result shows that the protein has toxicity to the varroa destructor, and under a condition that the activity of the chitinase ChiCl is 14.7 U / ml, the varroa destructor transfected by the protein has a mortality rate of 25 percent after 5 days. Sequencing analysis shows that the gene is a new sequence of chitinase gene ChiCl types. The invention can be used in an aspect of preventing the mite damage of bees.

Description

Technical field: [0001] The invention belongs to the field of biotechnology, and in particular relates to chitinase ChiC1 with activity of killing Varroa mite and its application in killing Varroa mite. Background technique: [0002] Bee parasitic mites have always been one of the most important hazards to the world's beekeeping industry. They are harmful on a global scale. At present, except for Australia and some parts of Africa, bee mites have not been found, and bee colonies in other continents are harmed by bee mites. The ectoparasitic mite Varroa destructor Anderson&Trueman (Varroa destructor Anderson&Trueman) sucks the body fluids of bee adults or larvae, causing the decline of bee physique and the weakening of bee colonies; in addition, bee mite also spreads bee bacteria and viruses disease. According to reports, the honeybee colony collapse disorder (CCD) that recently appeared in the United States and other countries is also related to the harm of bee mites. Ever...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N9/42A01N63/02A01P7/02C12R1/425
Inventor 韩日畴涂爽
Owner INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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