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Gene chip of inspection and quarantine fruit fly

A gene chip, fruit fly technology, applied in the field of molecular biology, can solve the problem that the detection fruit fly species cannot be quickly and accurately determined.

Inactive Publication Date: 2010-09-29
SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The present invention aims to solve the technical problem that the prior art cannot quickly and accurately determine the species of fruit flies

Method used

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  • Gene chip of inspection and quarantine fruit fly
  • Gene chip of inspection and quarantine fruit fly
  • Gene chip of inspection and quarantine fruit fly

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1 Determination of the fruit fly species to be detected

[0061] According to the representativeness of the main genus of Tephritidae, my country's list of quarantine pests, intergovernmental agreements, and domestic distribution, the present invention has determined that there are 20 species of quarantine fruit flies that are of great significance in my country. The categories included in these categories are described in Table 5.

[0062] Table 5 20 kinds of quarantine fruit flies targeted by the present invention

[0063] belongs to

Embodiment 2

[0064] Example 2 Determination and Analysis of the COI Sequence of Fruit Fly to be Detected

[0065] 1. Fruit fly DNA extraction

[0066] Species sample DNA can be extracted from a variety of tissues and organs, and factors such as the freshness, storage method, and storage time of the specimen may affect the extraction method and DNA quality. Fresh, unpreserved specimens are best, as once the specimen dies, cell lysis and the enzymes it releases will quickly degrade the DNA. Therefore, the fresher the specimen, the better.

[0067] According to the characteristics of fruit fly specimens, the high-salt method was used to extract DNA. The specific extraction steps are as follows:

[0068] (1) 500μl buffer (10mM Tris, 100mM NaCl, 10mM EDTA, 0.5-1% SDS) plus tissue (partial or whole fruit fly sample);

[0069] (2) Add 10 μl proteinase K (20mg / ml);

[0070] (3) Digestion at 55°C (digestion is subject to tissue dissolution, and the time is greater than 7 hours)

[0071] (4) A...

Embodiment 3

[0095] The determination of the specific oligonucleotide probe of the fruit fly to be detected in embodiment three

[0096] 1 probe design

[0097] Probe design should meet the one-to-one correspondence between the designed probe and the species, and in order to meet the conditions of chip hybridization, it is necessary to fully consider 1) the probe itself has no secondary structure; 2) all probes should maintain similar Tm values , GC content; 3) The selected probes are generally hypervariable regions in the sequence, so different haplotypes may also appear in the same species, so if necessary, use merger probes and other factors.

[0098] Due to the limitations of some of the above factors, the selected probes need to be verified repeatedly. Due to complex hybridization dynamics factors encountered in actual chip hybridization, the probes were adjusted repeatedly according to the actual hybridization situation. According to the COI sequence introduced above, the three pro...

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Abstract

The invention relates to the technical field of molecular biology, in particular to a gene chip of inspection and quarantine fruit fly. The invention aims to solve the technical problem that the existing technology can not determine the type of inspection fruit fly fast and accurately. The invention provides the gene chip of inspection and quarantine fruit fly so as to solve the technical problem. The gene chip comprises a solid phase carrier, wherein the oligonucleotide probes of an oligonucleotide probe set of inspection and quarantine fruit fly are fixed on the surface of the solid phase carrier, and the oligonucleotide probes can perform hybridization reaction with samples to be detected respectively. The gene chip of inspection and quarantine fruit fly has good application prospect in the detection and identification of quarantine fruit fly.

Description

technical field [0001] The invention relates to the technical field of molecular biology, in particular to a gene chip for detecting quarantine fruit flies by using oligonucleotide probes. Background technique [0002] Entry-exit plant inspection and quarantine is committed to preventing the invasion, colonization, and spread of plant dangerous sexually transmitted diseases, insects, weeds, and other harmful organisms, and is closely related to agricultural production safety and trade. In recent years, with the development of trade brought about by economic take-off and the increasing frequency of international exchanges, the chances of dangerous venereal diseases, insects, weeds and other harmful organisms being introduced into my country have also greatly increased; in addition, plant quarantine is an important non-tariff Therefore, it is becoming more and more obvious as a technical barrier to international trade. [0003] Fruit-fly plays an important role in plant inspec...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
Inventor 范京安曾晓茂陈世界莫邦辉温演庆夏云何万兴刘伟
Owner SICHUAN ENTRY EXIT INSPECTION & QUARANTINE BUREAU OF THE PEOPLES REPUBLIC OF CHINA
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