A gene chip for specific detection of Bacillus megaterium

A technology of Bacillus megaterium and specific genes, applied in the application field of Bacillus megaterium DNA detection chip and its preparation, microbial fertilizer and soil microbial detection, can solve the problems of poor specificity, small 16SDNA difference, classification and identification of Bacillus thuringiensis, etc. problem, achieve reliable results and provide specificity

Inactive Publication Date: 2019-09-24
山西刻选环境科技有限公司
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  • Application Information

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Problems solved by technology

At present, the traditional microbial detection method is mainly the culture method, which has a long detection cycle and cannot classify and identify closely related Bacillus megaterium, Bacillus subtilis, and Bacillus thuringiensis
In recent years, barcode sequences similar to 16S DNA have also been frequently used in the identification of microorganisms, but the difference between species of 16S DNA is very small, so it is inevitable to have poor specificity for classification and identification only by 16S DNA

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  • A gene chip for specific detection of Bacillus megaterium
  • A gene chip for specific detection of Bacillus megaterium
  • A gene chip for specific detection of Bacillus megaterium

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Embodiment Construction

[0022] Concrete implementation steps of the present invention are as follows:

[0023] Primers and probes were designed by our laboratory and synthesized by Shanghai Sangon Bioengineering Co., Ltd.

[0024] (1) Determination of the target sequence: According to the reported Bacillus megaterium 16SDNA sequence and rpoB gene sequence, select the Bacillus megaterium rpoB specific gene fragment whose nucleotide sequence is SEQ ID NO: 1, and the nucleotide sequence is SEQ ID NO: The 16S specific gene fragment of Bacillus megaterium 2, which is used as the target sequence for detection.

[0025] (2) Design of primers and probes: After the target sequence is determined, according to the design principles of primers and probes, design primers and probes as follows:

[0026] Bacillus megaterium rpoB:

[0027] Primer 1: 5’FAM—GTGTAATTTCACGTATTTTTACCG—3’

[0028] Primer 2: 5'-GCTCATGGTCCTCTTCTGAGTCC-3'

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Abstract

The invention discloses a gene chip for detecting the specificity of bacillus megatherium, and belongs to microorganism gene detection techniques. According to the reported 16SDNA sequence and an rpoB gene sequence of the bacillus megatherium, unique gene fragments rpoB and 16S of the bacillus megatherium are chosen as a detection target sequence. A corresponding multiplex PCR amplimer pair, a probe and a detection chip are also designed. Through the gene detection chip, the bacillus megatherium is qualitatively and quantitatively detected within a short period of time, and the classification and identification of the bacillus megatherium on a species level. The gene chip provides a powerful tool for tracking colonization, reproduction and the like of the bacillus megatherium in a fertilizer or soil.

Description

Technical field: [0001] The invention relates to a microorganism DNA detection technology, in particular to a bacillus megaterium DNA detection chip and its preparation, as well as its application in microbial fertilizer and soil microbial detection. [0002] technical background: [0003] Bacillus megaterium, a functional microorganism, is often used to make bio-organic fertilizers and in soil to regulate the microbial flora structure to achieve disease resistance, growth promotion, and quality improvement of crops. However, how to qualitatively and quantitatively track the colonization and reproduction behavior of this bacterium in fertilizers and soils has been perplexing relevant workers. The conventional detection of the number of effective viable bacteria has been unable to meet the needs of scientific research and production. The qualitative and quantitative analysis of common functional bacteria by gene chip technology has strong application value. At present, the tr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6837C12Q1/04C12N15/11C12R1/11
Inventor 任振兴王俊红王梦亮刘滇生
Owner 山西刻选环境科技有限公司
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