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Primers for rapidly identifying bemisia tabaci sodium ion channel gene mutation T929V and application thereof

A sodium ion channel and Bemisia tabaci technology, applied in the field of molecular biology, can solve problems such as the design of specific primers that cannot be fully applied, and achieve the effect of improving the accuracy rate

Inactive Publication Date: 2017-01-04
QINGDAO AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although this technical scheme introduces a mismatched base G to the third base at the 3' end of the mutant forward primer, thereby improving the specificity of the primer, this method is not fully applicable to all B. tabaci sodium channels Design of specific primers for gene mutation sites

Method used

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  • Primers for rapidly identifying bemisia tabaci sodium ion channel gene mutation T929V and application thereof
  • Primers for rapidly identifying bemisia tabaci sodium ion channel gene mutation T929V and application thereof
  • Primers for rapidly identifying bemisia tabaci sodium ion channel gene mutation T929V and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0043] Sequencing verification of wild-type and mutant templates of sodium ion channel gene T929V in Bemis tabaci

[0044] Design specific primers according to the Bemisia tabaci sodium ion channel gene (Genbank: DQ205209.1) published by NCBI, perform PCR amplification on the 929-position mutation, and directly perform PCR sequencing after recovering the specific target fragment.

[0045] Primers for PCR amplification:

[0046] Upstream primer: GGCCAACTTTGAATCTGTT;

[0047] Downstream primer: AAACTTTCCGCACCTCTG.

[0048] PCR amplification system: Genomic DNA solution 2μl, 10M upstream primer 1μl, 10M downstream primer 1μl, 5U / μl rTaq enzyme 0.25μl, 10×Taq Buffer (buffer) 2.5μl, 10mM dNTP 2μl, ddH 2 0 to 25 μl;

[0049] PCR amplification conditions: pre-denaturation at 94°C for 3 minutes; denaturation at 94°C for 30 seconds, annealing at 50°C for 30 seconds, extension at 72°C for 30 seconds, and 35 cycles; extension at 72°C for 7 minutes.

[0050] According to the above, th...

Embodiment 2

[0065] A primer for quickly identifying the mutation site T929V of the sodium ion channel gene as wild-type Bemisia tabaci, the primers are a pair, the nucleotide sequence of the upstream primer is shown in SEQ ID NO.1, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.1 Shown in ID NO.2.

[0066] A primer for quickly identifying the mutation site T929V of the sodium ion channel gene as a mutant whitefly, the primers are a pair, the nucleotide sequence of the upstream primer is shown in SEQ ID NO.3, and the nucleotide sequence of the downstream primer is shown in SEQ ID NO.3 Shown in ID NO.2.

[0067] The method for identifying the T929V genotype of the Bemisia tabaci sodium ion channel gene mutation site using the above primers, the steps are as follows:

[0068] (1) extracting the genomic DNA of the sample to be identified to obtain a genomic DNA solution;

[0069] (2) Using the genomic DNA obtained in step (1) as a template, use the primers of the...

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Abstract

The invention relates to primers for rapidly identifying bemisia tabaci sodium ion channel gene mutation T929V and application thereof. According to primers for rapidly identifying a sodium ion channel gene mutation site T929V as wild bemisia tabaci, the upstream primer nucleotide sequence is shown in SEQ ID NO.1, and the downstream primer nucleotide sequence is shown in SEQ ID NO.2. According to the primers for rapidly identifying the sodium ion channel gene mutation site T929V as mutant bemisia tabaci, the upstream primer nucleotide sequence is shown in SEQ ID NO.3, and the downstream primer nucleotide sequence is shown in SEQ ID NO.2. The two pairs of primers are adopted for detection, detection accuracy can be improved, the occurrence and development trend of drug resistance of field bemisia tabaci to pyrethroid insecticide can be rapidly and sensitively detected, a bemisia tabaci preventing and treating strategy can be adjusted in real time to delay and control development of drug resistance.

Description

technical field [0001] The present invention relates to a primer for quickly identifying the mutation T929V of the Bemisia tabaci (Gennadius) Para sodium ion channel gene and its application, in particular to a primer for identifying the mutation T929V of the Para sodium ion channel gene in the whitefly Bemisia tabaci (Gennadius) and its application in pyrethroids The application in the monitoring of insecticide resistance belongs to the technical field of molecular biology. Background technique [0002] Bemisia tabaci B.tabaci is an important field and greenhouse pest, which can cause damage to more than 600 kinds of plants through direct feeding, honeydew secretion and plant virus transmission. Due to its rapid adaptability, Bemisia tabaci has developed a high level of resistance to various insecticides including organophosphates and carbamates. It is the only pest dubbed the title of "super pest" by the scientific and technological circles. [0003] Pyrethroid insecticid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11C12N15/10
CPCC12Q1/6888C12Q1/6806C12Q2600/124C12Q2600/156
Inventor 郭磊褚栋李增鑫房宽
Owner QINGDAO AGRI UNIV
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