Detection method for identifying amycolatopsis

A technology of amycobacteria pseudomycoides and a detection method, which is applied in the field of primer pairs and detection for the detection of Amycolatopsis pseudomycoides, and can solve problems such as bacterial species identification that is unfavorable for a large sample size, lack of rapid detection methods, and long time consumption. , to achieve the effect of fast method, short time-consuming and good effect

Active Publication Date: 2021-06-04
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Claims
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Problems solved by technology

However, although the above method can identify pseudoabranchic acid, it is costly and time-consuming.
[0005] At present, there are no specific amplification primers for Amycolatopsis genus, and the lack of rapid detection methods is not conducive to the identification of large sample sizes

Method used

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  • Detection method for identifying amycolatopsis
  • Detection method for identifying amycolatopsis
  • Detection method for identifying amycolatopsis

Examples

Experimental program
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Embodiment 1

[0033] 1. Primer synthesis

[0034] Analyze the AMETH_3452 gene and its homologous sequences, and design PCR primers, see Table 1 for details. The primers were synthesized by Sangon Bioengineering (Shanghai) Co., Ltd. according to the sequence in the sequence listing.

[0035] Table 1

[0036]

[0037]

[0038] In the table, S=C or G; R=A or G; N=A, C, G or T; Y=C or T.

[0039] 2. Optimization of PCR reaction conditions

[0040] 2.1 The environmental wastewater DNA was used as a blank control, and the known DNA of A. methanolica 239 was used as a positive control. The bacterial strains were isolated and cultured, and the mycelium was directly scraped from the cultured petri dish, and the genomic DNA of the mycelium was extracted with a bacterial DNA extraction kit (Shanghai Jierui Bioengineering Co., Ltd.).

[0041] 2.2 Use the extracted DNA as a template, use the above primers, and use PCR to amplify. The annealing temperature of PCR was optimized by gradient PCR me...

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Abstract

The invention discloses a detection method for identifying amycolatopsis, and belongs to the field of molecular biology. The invention provides a primer pair capable of specifically identifying the amycolatopsis. The primer pair is designed based on a specific conserved gene AMETH_3452 in the amycolatopsis and homologous gene segments in the amycolatopsis and has strong specificity; a PCR method for rapidly detecting the amycolatopsis is established through optimization of PCR parameters, and it is repeatedly proved that the method is rapid and sensitive, a plurality of strains can be detected at the same time, the detection cost is low, consumed time is short, and the effect is good. A basis is provided for further research on environment detection and animal or clinical actinomycetes.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular to a pair of primers and a detection method for detecting the genus Amycolatopsis. Background technique [0002] The secondary metabolites produced by microorganisms have various biological activities and are currently an important treasure trove for drug discovery. Microbial drugs have the characteristics of rich sources and unique chemical structures of most secondary metabolites produced by microorganisms, which are difficult to synthesize. Drugs have important clinical application value. [0003] Amycolatopsis (Amycolatopsis) is an important class of antibiotic-producing actinomycetes, such as vancomycin-producing bacteria Amycolatopsis orientalis (Amycolatopsis orientalis) and rifamycin-producing bacteria Amycolatopsis mediterranei ( Amycolatopsis mediterranei). Most of the strains of Amycolatopsis are isolated from soil, and some are isolated from oceans, deserts, sediments...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/689C12Q1/686C12Q1/04C12N15/11C12R1/01
CPCC12Q1/689C12Q1/686C12Q2565/125
Inventor 唐标杨华吴静钱鸣蓉
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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