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Amplification primers for the full-length sequence of the coiii gene of Ixodes tick and its application

A technology of COIII-TF and COIII-TR, which is applied in the field of molecular biology, can solve the problems of lack of general primers for amplifying Ixodidae COⅢ gene, few Ixodes COⅢ sequences, and restrictions on the application of COⅢ sequences

Active Publication Date: 2018-12-21
CHINESE ACAD OF INSPECTION & QUARANTINE
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Problems solved by technology

However, there are few Ixodes COⅢ sequences in the Genbank database, and the lack of general primers for amplifying the Ixodidae COⅢ gene severely restricts the application of COⅢ sequences in tick phylogeny and tick-borne epidemiological investigations.

Method used

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  • Amplification primers for the full-length sequence of the coiii gene of Ixodes tick and its application
  • Amplification primers for the full-length sequence of the coiii gene of Ixodes tick and its application
  • Amplification primers for the full-length sequence of the coiii gene of Ixodes tick and its application

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Embodiment 1

[0024] 1. Data collection: Download the mitochondrial COⅢ gene sequences of different species of Ixodidae from Genbank (NCBI, http: / / www.ncbi.nlm.nih.gov / )

[0025] 2. Sequence comparison: use the ClustalW program in the MEGA software to compare the collected COⅢ sequences of Ixoderidae ticks, and find out the sequence fragments that are more conservative and have the least base variation.

[0026] 3. Primer synthesis: According to the comparison results above, use the primer design software Primer5 to design a pair of merger amplification primers in the sequence fragment with the smallest base variation.

[0027] The primer sequences are:

[0028] COIII-TF: 5'-ATGATATTTCATCCTTTTCACTTAGT-3';

[0029] COIII-TR: 5'-AATAAATTCATCAATATATAAATGTA-3'.

[0030] 4. Primer amplification: The above primers were respectively applied to the PCR amplification of COⅢ sequences collected from 265 individuals of 11 species of 5 genera of Ixodidae shown in Table 1.

[0031] Table 1 11 species...

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Abstract

The invention relates to the field of molecular biology, and particularly provides amplification primers of an ixodidae tick COIII full-length gene sequence and application thereof. The amplification primers of the ixodidae tick COIII full-length gene sequence are shown in SEQ ID No.1 and SEQ ID No.2. The ixodidae mitochondrion COIII gene sequence full length (778bp) can be effectively amplified, and the amplification efficiency reaches 98.5 percent.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to an amplification primer for the full-length sequence of the COIII gene of Ixoderidae ticks and an application thereof. Background technique [0002] The family Ixodidae belongs to the phylum Arthropoda, the class Arachnida, and the order Ixodida. At present, there are more than 700 species of hard ticks known in the world, and the effective species of hard ticks recorded in my country belong to the genus Ixodes, Amblyomma, Haemaphysalis, Dermacentor, Glass There are 106 species in 7 genera of Hyalomma, Rhipicephalus and Anomalohimalaya. Ticks suck blood from their hosts and are very harmful to humans and domestic animals. The bite of ticks not only causes blood loss to the host, but also causes symptoms such as inflammation and ulcers on the host's skin. More importantly, as a medium, hard ticks can transmit human and animal diseases, including viral diseases (forest encephalit...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12Q1/6888
Inventor 吕继洲韩雪清王慧煜袁向芬贾广乐
Owner CHINESE ACAD OF INSPECTION & QUARANTINE
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