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Gene chip for performing specific detection on bacillus subtilis

A technology of Bacillus subtilis and specific genes, which is applied in genetic engineering, plant genetic improvement, microbial measurement/inspection, etc., can solve the problems of 16SDNA small difference, poor specificity, classification and identification of Bacillus thuringiensis, etc., and provide specificity , reliable results

Inactive Publication Date: 2016-12-07
SHANXI UNIV
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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 subtilis, 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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  • Gene chip for performing specific detection on bacillus subtilis
  • Gene chip for performing specific detection on bacillus subtilis
  • Gene chip for performing specific detection on bacillus subtilis

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Embodiment Construction

[0021] Concrete implementation steps of the present invention are as follows:

[0022] Primers and probes were designed by our laboratory and synthesized by Shanghai Sangon Bioengineering Co., Ltd.

[0023] (1) Determination of the target sequence: According to the reported Bacillus subtilis 16SDNA sequence and rpoB gene sequence, select the Bacillus subtilis rpoB specific gene fragment whose nucleotide sequence is SEQ ID NO: 1, and the nucleotide sequence is SEQ ID NO: 2 Bacillus subtilis 16S specific gene fragment, which is used as the target sequence for detection.

[0024] (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:

[0025] Bacillus subtilis rpoB:

[0026] Primer 1: 5'FAM-GCTTCGCAATCTTGATGACCGC-3';

[0027] Primer 2: 5'-GTAAGGGCCTGTAGAGCGTGCA-3';

[0028] 5'NH 3 —TTTTTTTTTTTAACTGAACTGACTGCCGAAGAACGCC-3'.

[0029] Bacillus subtilis 16S:

[0...

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Abstract

The invention discloses a gene chip for performing specific detection on bacillus subtilis, and belongs to soil microbe gene detection technology. According to a reported bacillus subtilis 16SDNA sequence and a reported rpoB gene sequence, a bacillus subtilis rpoB-specific gene fragment and a 16S-specific gene fragment are selected as detected target sequences, and corresponding multiple PCR amplification primer pairs, probes and detection chips are designed. Through the gene detection chip, the bacillus subtilis can be qualitatively and quantitatively detected within a short time, and classification and identification of the bacillus subtilis in the species level are achieved. The gene chip provides a powerful tool for colonization and propagation of the bacillus subtilis in fertilizer or soil and the like.

Description

Technical field: [0001] The invention relates to a microorganism DNA detection technology, in particular to a bacillus subtilis DNA detection chip and its preparation, as well as its application in soil microorganism detection. technical background: [0002] Bacillus subtilis, a functional microorganism, is often used to make bio-organic fertilizers and in soil to regulate the microflora 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 traditional microbial detection method ...

Claims

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Application Information

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IPC IPC(8): C12N15/31C12N15/11C12Q1/68C12Q1/04
CPCC12Q1/6837C12Q1/689C12Q2600/16C12Q2537/143C12Q2545/113C12Q2565/501
Inventor 任振兴王俊红王梦亮刘滇生
Owner SHANXI UNIV
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