Primer sequence for detecting tetracycline resistant gene tetO in water body and sediments in classified mode and corresponding detecting method

Abstract

The invention provides a primer sequence for detecting a tetracycline resistant gene tetO in a water body and sediments in a classified mode and a corresponding detecting method for detecting the tetracycline resistant gene tetO in the water body and the sediments through the primer sequence. The primer sequence comprises a tetO-F, a tetO-R and a tetO-F-GC. The PCR amplification and denaturing gradient gel electrophoresis technology are conducted through the primer sequence, and then the aim of detecting the tetO sequence in the classified mode is achieved. By designing a primer and applying the PCR-DGGE technology to the classified identification of the tetO sequence, the steps, highly consuming labor, time and expenses, of the separation and purification of DNA, the connection and conversion of carriers, the screening of positive clone, the large-scale sequencing of sequences and the like are omitted compared with a traditional method, the comprehensive tetO sequence information in a sample can be obtained, the whole operation becomes easy and rapid, and cost is saved.

Description

technical field [0001] The invention relates to the fields of DNA extraction technology, PCR amplification technology, denaturing gradient gel electrophoresis technology and traditional sequencing technology in water bodies and sediments, and more specifically relates to a method for classifying and detecting tetracycline resistance gene tetO in water bodies and sediments. Primer sequences and corresponding detection methods, in order to finally achieve the purpose of separating and identifying different tetO sequences. Background technique [0002] Antibiotics are microbial secondary metabolites that can selectively inhibit or kill other microorganisms or tumor cells at low concentrations, as well as similar compounds and structural modifications prepared by chemical or biological methods. Antibiotics are widely used in the pharmaceutical industry and livestock and poultry breeding industry. In recent years, the domestic and iatrogenic use of antibiotics has increased sign...

AI Technical Summary

Problems solved by technology

However, for the current tetO gene detection technology, there are conventional qualitative PCR methods and quantitative PCR methods based on fluorescent dyes. However, when there are multiple tetO genes mixed in the sample, the above methods cannot effectively detect multiple tetO genes. Effectively classify and identify

Although the traditional method of amplifying genes into carrier bacteria through ligation transformation, and then selecting a large number of positive clones through blue-white screening and sequencing can achieve the purpose of comprehensively analyzing different genes in samples, the operation is time-consuming and laborious, and the post-analysis costs are relatively high. high, so not commonly used

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