Chitinase ChiCl for deactivating varroa destructor and application thereof

A technology for killing Varroa dissociates and chitinase, which is applied in the fields of application, acaricides, enzymes, etc., can solve the problem that no chitinase has been found to control Varroa disses in bees

Inactive Publication Date: 2009-12-30
INST OF ZOOLOGY GUANGDONG ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, so far, no reports have been found on the use

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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

[0022] Serratia marcescens GEI of the present invention is isolated from the midgut of bee worker bees in Conghua by the inventors. This bacterium was preserved in the China Center for Type Culture Collection (CCTCC, Address: Wuhan University, Wuhan, China), deposit number CCTCC M 209097.

[0023] 1. Cloning and sequencing of the coding gene of chitinase ChiC1.

[0024] 1. Culture of Serratia marcescens GEI

[0025] Pick a single colony of Serratia marcescens GEI from the LB plate and inoculate it into 4 mL of LB liquid medium, and culture overnight with conventional shaking (28°C, 200r / min); 100mL Erlenmeyer flask with 10mL culture medium, cultivate to OD620≈2.

[0026] 2. Extraction of Bacterial Genomic DNA

[0027] Genomic DNA of Serratia marcescens strain GEI was extracted using a conventional kit (TIANGEN Bacterial Genome Extraction Kit, Beijing TIANGEN Company).

[0028] 3. Primer Design

[0029] According to the chitinase gene of Serratia marcescens which has been ...

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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. Varroa destructor Anderson & Trueman, the ectoparasitic mite of honeybees (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 viral diseases. 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 o...

Claims

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Application Information

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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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