Rhodococcus strain and identification and application methods thereof

A technology of Rhodococcus and strains, applied in the field of microorganisms, can solve problems such as long cycle

Active Publication Date: 2019-03-05
CHINA PETROLEUM & CHEM CORP +1
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  • Abstract
  • Description
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  • Application Information

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Problems solved by technology

[0004] At present, there are two common detection and identification methods for Rhodococcus: 1. Physiological and biochemical identification, through the isolated pure bacteria, qualitative research on the physiology and biochemistry of microorganisms, this method has a long cycle and puts forward higher requirements for the purity of the culture; 2. .After gene extraction, 16S rDNA sequencing is performed. This method still needs to obtain pure cultures for whole gene extraction, and then use general 16S rDNA primers to obtain conserved sequences for sequencing

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  • Rhodococcus strain and identification and application methods thereof
  • Rhodococcus strain and identification and application methods thereof
  • Rhodococcus strain and identification and application methods thereof

Examples

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Embodiment 1

[0069] Example 1: Using Rhodococcus SINOPEC42 to identify soil samples above oil and gas reservoirs.

[0070] (1) Design specific primers based on the conserved 16S rDNA sequence of Rhodococcus SINOPEC42. The sequences of the designed specific primers are as follows:

[0071] Upstream primer: 5'-AGCTTGCAGGCGATACGGGCAGACTTGA-3';

[0072] Downstream primer: 5'-CGCTCGTTGCGGGACTTAACCCAACATC-3';

[0073] The size of the target band amplified by the specific primer is about 480bp.

[0074] (2) Extract the whole genome of microorganisms in the soil.

[0075] Weigh 0.5g of the soil above the oil and gas reservoir and the background soil, add them to the Lysing Matrix E Tube respectively, and use a cell disruptor to break the cells (speed 4.5m / s, 30s, 4 times), and the broken cells are according to SPINKit for Soil (U.S. MP Biomedicals biomedical company product) instruction manual Carry out follow-up operation, obtain the complete genome of microorganism in the soil sample, as th...

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Abstract

The invention relates to a rhodococcus strain and identification and application methods thereof in the technical field of microorganisms. The rhodococcus strain has a 16S rDNA (16S ribosome deoxyribonucleic acid) gene sequence shown as SEQ ID NO.1, a strain name of SINOPEC42 and a preservation number of CCTCC NO: M2016112 in China Center for Type Culture Collection. The abundance of the rhodococcus strain in soil above an oil reservoir is positively related to the concentration of floating gaseous hydrocarbons of the oil reservoir, so that the rhodococcus strain can serve as an oil exploitation microorganism to mark areas with high hydrocarbon leakage above the oil reservoir; meanwhile, the rhodococcus strain is high in abundance in soil above the oil reservoir and saves amplification culture, the 16S rDNA conserved sequence of the rhodococcus strain SINOPEC42 in soil through improved primers, so that the concentration of floating gaseous hydrocarbons of the oil reservoir can be determined accurately and efficiently, and the deficiency of long detecting cycle and high requirements on purity of sample culture in traditional physiological and biochemical detection can be solved.

Description

technical field [0001] The invention belongs to the technical field of microorganisms, and in particular relates to a rhodococcus and its identification method and application. Background technique [0002] The core of microbial exploration technology is the identification of oil and gas indicator microorganisms. Usually, the relative abundance of the target microorganisms studied in the exploration process is very low in the environment, so it is necessary to stimulate the rapid reproduction of hydrocarbon oxidizing bacteria in the oil and gas reservoir environment and non-oil and gas reservoir environment under the traditional specific culture conditions. Although the above method can obtain the quantity information of microorganisms, the quantity information is actually geometrically higher than the number of microorganisms in the in-situ environment, which inevitably conceals the difference in the number of microorganisms in oil and gas reservoirs and non-oil and gas res...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12Q1/04C12R1/01
CPCC12Q1/04C12Q1/689C12R2001/01C12N1/205Y02W10/10
Inventor 汤玉平赵克斌金之钧顾磊许科伟杨帆孙长青
Owner CHINA PETROLEUM & CHEM CORP
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