Radiation resistant microbacterium and use thereof

A technology of radiation resistance and microbacteria, which is applied in the field of microorganisms, can solve the problems that the research on radiation-resistant microorganisms started late, and there are no new species or genera of extremely radiation-resistant microorganisms, and achieve the effect of strong resistance to ultraviolet radiation

Active Publication Date: 2009-07-15
GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There have been more than 20 years of research and development of microbial resources in my country, but no new species or genera of extremely radiation-resistant microorganisms have been reported, and the research on related radiation-resistant microorganisms started relatively late

Method used

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  • Radiation resistant microbacterium and use thereof
  • Radiation resistant microbacterium and use thereof
  • Radiation resistant microbacterium and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Excavate soil samples from the soil in Xinjiang nuclear radiation areas, take 10g of soil and put it into a triangular flask (with glass beads) filled with 90mL sterile water, shake it at 180rpm for 30 minutes, let it stand for 10 minutes, and take the upper layer to dilute After 10 times, take 0.1mL and spread it evenly on the nutrient agar medium plate, turn the plate over and place it in a 28°C incubator and cultivate it for 2 days to obtain Microbacterium radiodurans GIMN1.002.

[0023] Medium: Use nutrient agar medium. The specific formula is: peptone 10.0g, yeast extract 5g, sodium chloride (NaCl) 5g, agar 20.0g, water 1000ml, pH6.8~7.0;

[0024] Culture temperature: 28°C.

[0025] Radiation-resistant Microbacterium Microbacterium radiodurans GIMN1.002 of the present invention has the following microbial characteristics:

[0026] 1. Morphological characteristics

[0027] The strain GIMN1.002 has small colonies on the plate, the surface is smooth and translucent...

Embodiment 2

[0039] Embodiment 2: 16SrRNA gene sequence analysis

[0040] 1. Rapid preparation of PCR template DNA

[0041] The bacterial strain obtained in Example 1 was streak-inoculated on a plate of NA medium, and cultured overnight at 30°C. Suspend a single colony in 10 μl sterile distilled water, heat in boiling water for 5 min, centrifuge, and take the supernatant as PCR template DNA.

[0042] 2. PCR amplification of 16SrRNA gene sequence

[0043] PCR primers were synthesized by Shanghai Yingjun Company.

[0044] PrimerA: 5'-AGAGTTTGATCCTGGCTCAG-3'

[0045] Primer B: 5'-AAGGAGGTGATCCACCCCCA-3'

[0046] PCR reaction system (total volume 50 μl): 5 μl of 10x buffer, 4 μl of dNTP (2 mmol / l), 1 μl of PrimerA (10 pmol / l), 1 μl of PrimerB (10 pmol / l), 0.4 μl of Taq enzyme (2U / l), DNA Template 1 μl, sterilized ddH 2 O 37.4 μl.

[0047] PCR amplification conditions: 94°C for 5 min; 94°C for 1 min, 58°C for 1 min, 72°C for 2 min, 30 cycles; 72°C for 7 min.

[0048] 3. Sequence determina...

Embodiment 3

[0050] Embodiment 3: Determination of the resistance to ultraviolet radiation of the radiotolerant microbacteria of the present invention

[0051] Inoculate one ring of the radioresistant Microbacterium radiostrain of the present invention into a liquid nutrient medium (NA medium without agar), culture on a shaker until the logarithmic phase of growth, and collect the bacteria by centrifuging at 5000 rpm for 10 minutes at 4°C; Bacteria were resuspended in an equal volume of 0.1M potassium phosphate buffer (pH 7.0) (K 2 HPO 4 27.816g, KH 2 PO 4 10.608g, dissolved in 1L of distilled water, sterilized at 121°C for 20min), then collected by centrifugation at 5000rpm at 4°C for 10min, and resuspended in an equal volume of 0.1M potassium phosphate buffer (pH 7.0).

[0052] Place the bacterial solution in a Petri dish and stir it with a sterilized iron wire to make the bacterial cells uniform. Place the petri dishes at 0~500J / m 2 Under different doses of ultraviolet irradiatio...

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Abstract

The invention discloses a radioresistant Microbacterium radiodurans GIMN1.002 and uses thereof, the bacterium has been collected in China Center for Type Culture Collection, the collection date is 11.10.2008 and the collection number is CCTCC No.M208212. The ability of the inventive radioresistant Microbacterium radiodurans to resist the ultraviolet radiation is significant for researching on repair molecular mechanism of DNA damage thereof and for promoting the development of new DNA technologies in environmental protection, biomediation, human health, etc.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to a new bacterium capable of enduring ultraviolet radiation and its application. Background technique [0002] Since American scientist Anderson isolated Deinococcus radiodurans from canned meat that deteriorated after radiation sterilization in 1956, it has attracted the attention of microbiologists, radiation biologists and tumor researchers. The development and research of such microbial strain resources and their ability to repair DNA damage are of great significance not only to the basic science of exploring the molecular mechanism of DNA repair, but also to promoting the development of new DNA technology and in environmental protection, bioremediation, and human health. , biotechnology, and even the development and utilization of extraterrestrial space have great application prospects. There have been more than 20 years of research and development of microbial resources in ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12R1/01
Inventor 朱红惠张维王燕陈明姚青羊宋贞袁梦龙林敏黎志坤
Owner GUANGDONG INST OF MICROBIOLOGY GUANGDONG DETECTION CENT OF MICROBIOLOGY
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