A DNA bar code standard gene sequence for Taiwan Lasiohelea

A technology of gene sequence and barcode, applied in the field of species identification, can solve the problem of lack of gene sequence of the Taiwan midge, and achieve the effect of shortening the identification time.

Active Publication Date: 2012-09-26
FUJIAN INT TRAVEL HEALTH CARE CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the application of this method in the molecular identification of blood-sucking midges is limited to Culicoides. Genbank also has some COI gene sequences of Culicoides, but there is no related gene sequence of Taiwan Midges.

Method used

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  • A DNA bar code standard gene sequence for Taiwan Lasiohelea
  • A DNA bar code standard gene sequence for Taiwan Lasiohelea
  • A DNA bar code standard gene sequence for Taiwan Lasiohelea

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] 1. Collection and preservation of specimens of midges in Taiwan

[0034] The algal midges were collected by human trap or human trap, and the collected specimens were killed by freezing and stored in 75% ethanol directly.

[0035] 2. Pretreatment of test samples

[0036] Take out the specimens of Midges taiwanensis preserved in 75% alcohol, and cut off the head, genitals and wings with a dissecting needle, which is convenient for making slices for species identification. The remaining part was transferred into PCR tubes, preserved in 75% alcohol, one for each tube, and uniquely numbered for DNA extraction. If it needs to be stored for a long time, the PCR tube containing the remaining tissue parts of C. formosanus should be stored in a -20°C refrigerator.

[0037] 3. DNA template preparation

[0038] Using the micro-DNA template preparation method of small insects to extract the DNA of midges in Taiwan (Editor-in-Chief Wen Lizhang. Introduction to Entomological Rese...

Embodiment 2

[0056] 1. Selection and storage of samples

[0057] Formosan midges bred in batches in the laboratory were killed by freezing and stored in 75% ethanol directly. The species of Taiwan midges have been reviewed by authoritative experts before batch breeding.

[0058] 2. Pretreatment of samples

[0059] Take out the specimens of Midges taiwanensis preserved in 75% alcohol, cut off the head and the end of the abdomen with a dissecting needle, transfer them into PCR tubes, store them in 75% alcohol, and carry out a unique number for DNA extraction.

[0060] 3. DNA template preparation

[0061] Using the micro-DNA template preparation method of small insects to extract the DNA of midges in Taiwan (Editor-in-Chief Wen Lizhang. Introduction to Entomological Research Methods and Techniques [M]. Beijing: Science Press, 2010.278-286), the extracted DNA samples were stored at -20°C spare.

[0062] 4. Primer synthesis

[0063] The primers used in this example are as follows:

[006...

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Abstract

The invention discloses a DNA bar code standard gene sequence for Taiwan Lasiohelea, which is characterized in that a bar code standard detection gene is COI gene, possessing gene sequence SEQ ID NO.1. Manual proofreading and sequence assembly are carried out to the sequencing result and Blast similarity search is carried out in NCBI to ensure that the obtained sequence is the target sequence. Species identification of Taiwan Lasiohelea is realized through morphological identification, thereby result reliability is guaranteed. The DNA bar code standard gene sequence for Taiwan Lasiohelea provided in the invention is in favor of realizing rapid identification of Taiwan Lasiohelea, and reduces identification time.

Description

technical field [0001] The invention relates to the field of species identification, in particular to a standard gene sequence of the DNA barcode of the Taiwan midge midge. Background technique [0002] Taiwan midge ( Lasiohelea taiwana Shiraki, 1913) commonly known as the small black mosquito, is a kind of tiny blood-sucking midges common in southern my country, belonging to the genus Ceratopogonidae (Ceratopogonidae) of the order Diptera ( Lasiohelea ). The midge bites the exposed places such as the calf and ankle of the human body during the day, causing itching and even an allergic reaction to the human body. Scholars in my country have successively isolated JE virus from the midge in Taiwan, and proposed that the midge may be a new transmission medium of JE virus. [0003] The identification and recognition of blood-sucking midges is the basis for monitoring and controlling the occurrence of midge-borne diseases, and it is also the core step in the field of disease...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/12C12N15/11C12Q1/68
Inventor 黄恩炯肖武王光辉张建庆王飞鹏方义亮房长天史灵梅
Owner FUJIAN INT TRAVEL HEALTH CARE CENT
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