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Induced mutation screening method and applications of Rhodococcus erythropolis bacterial strain YF28-1-4

A technique of mutagenesis screening and rhodococcus, applied in the biological field, can solve problems such as small-scale excision and frame-shift mutation, and achieve the effects of short cycle, simple operation, and high-efficiency crude oil degradation ability

Inactive Publication Date: 2018-07-06
COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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Problems solved by technology

NTG-induced mutations are mainly GC-AT transitions, in addition to small excisions, frameshift mutations, and loss of GC pairs

Method used

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  • Induced mutation screening method and applications of Rhodococcus erythropolis bacterial strain YF28-1-4
  • Induced mutation screening method and applications of Rhodococcus erythropolis bacterial strain YF28-1-4
  • Induced mutation screening method and applications of Rhodococcus erythropolis bacterial strain YF28-1-4

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Embodiment 1

[0026] The present invention as figure 1 The process flow chart shows the following steps,

[0027] Mutagenesis of Rhodococcus by heavy ion irradiation:

[0028] The treatment method of bacteria irradiated with heavy ions:

[0029] 1. Preparation of the medium and the solution used:

[0030] LB (1L): tryptone 10.0g, yeast extract 5.0g, sodium chloride 10.0g, distilled water to 1000 ml, pH adjusted to about 7.6.

[0031] pH7.0 phosphate buffer: K2HPO4 2g / L, KH2PO4 8 g / L, sterilized at 0.1 MPa for 20min.

[0032] MM medium: magnesium chloride 3.5g, ammonium nitrate 1.0g, potassium chloride 0.35g, calcium chloride 0.05g, potassium bromide 0.08g, ferric chloride 0.01g, metal salt solution 1mL (zinc sulfate 0.01%, strontium chloride 2.4%), phosphate solution 10mL (dipotassium hydrogen phosphate 10%, potassium dihydrogen phosphate 10%), agarose 15g, ddH2O to 1L, pH adjusted to about 7.5, finally add 1% (v / v) crude oil .

[0033] 2. Strain preparation:

[0034]The activated st...

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Abstract

The invention discloses an induced mutation screening method and applications of Rhodococcus erythropolis bacterial strain YF28-1-4. The induced mutation screening method comprises following steps: firstly, heavy ion irradiation is adopted for primary induced mutation of the genome of Rhodococcus erythropolis YF28-1(8) so as to obtain a bacterial strain with excellent degrading effect; then superpower mutagen NTG is adopted for a second time of induced mutation so as to obtain a Rhodococcus erythropolis mutant library, and then crude oil flat and GC-MS screening are adopted so as to obtain the Rhodococcus erythropolis bacterial strain YF28-1-4 with transcription system mutation and translation system mutation. The Rhodococcus erythropolis bacterial strain YF28-1-4 possesses high efficiency crude oil degradation capacity; operation of the induced mutation screening method is simple; period is short; efficiency is high; the induced mutation screening method is suitable for Rhodococcus erythropolis and other microorganisms with certain crude oil degrading capacity; and the popularization and application value is high.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for mutagenesis and rapid acquisition of highly efficient crude oil degradation bacteria and its application. Background technique [0002] As an important energy material and industrial raw material, crude oil and its by-products are being widely exploited by people all over the world, and they have a very wide range of roles and functions in social and economic development. However, due to limited technology and weak awareness of environmental protection, crude oil pollution in the environment has become increasingly serious, destroying the normal ecological balance. From 1996 to 1999 alone, there were about 407 oil spill accidents in Alaska every year; the Dalian Xingang oil spill incident in 2010 polluted at least 50 square kilometers of nearby sea area, with an impact area of ​​100 square kilometers. About 8 million tons of oil enter the environment every y...

Claims

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Application Information

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IPC IPC(8): C12N13/00C12N15/01A62D3/02C12R1/01A62D101/20
CPCA62D3/02A62D2101/20C12N13/00C12N15/01
Inventor 杨蕊琪常思静张威章高森陈拓刘光琇
Owner COLD & ARID REGIONS ENVIRONMENTAL & ENG RES INST CHINESE
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