Fusion method for breeding high-efficiency cadmium-resistant microbial protoplast multi-parent strains

A protoplast, multi-parent technology for use in microorganism-based methods, microbial assays/tests, biochemical equipment and methods, etc.

Inactive Publication Date: 2011-01-26
NANKAI UNIV
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Problems solved by technology

However, in terms of microbial treatment of heavy metal pollutants,

Method used

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Embodiment Construction

[0028] In order to have a sufficiently clear understanding of the present invention. The present invention will now be further described with reference to the embodiments.

[0029] The first step: primary mutagenesis of cultured strains

[0030] Bacillus subtilis or Candida tropicalis was inoculated into the slant medium, and after culturing at 30°C for a period of time, an appropriate amount of phosphate buffer was added to prepare suspended spores, so that the spore volume was 1 × 10 6 pcs / mL.

[0031] Step 2: Strain Mutagenesis

[0032] The mutagenesis treatment is as follows: take 5 mL of Candida tropicalis spore suspension and transfer it to a sterilized petri dish, and irradiate it under a 15W ultraviolet lamp for 5 min; take 5 mL of Bacillus subtilis spore suspension and irradiate for 1 min and 5 min; take 5 mL of Candida tropicalis The bacterial spore suspension was treated with a nitrous acid solution with a final concentration of 0.1 mol / L for 10 min; 5 mL of the ...

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Abstract

The invention discloses a method for improving cadmium tolerance of bacillus through mutation, which comprises the following steps of: (1) taking bacillus subtilis as a starting bacterium, and pre-culturing the bacillus on a beef extract peptone culture medium for 10 hours; and (2) performing ultraviolet mutation on pre-cultured active colony: irradiating by using 15W ultraviolet lamp under the condition of a fixed distance of 30cm for 5 minutes, performing mutation, screening by fermenting on a concentration gradient culture medium plate and in a shake flask, wherein the obtained active colony is the mutated bacillus with high tolerance to cadmium. Through the method, the mutated bacillus B38 fully tolerating high concentrate cadmium is obtained, and the cadmium tolerant concentration is 3mmol/L.

Description

technical field [0001] The invention relates to a microbial breeding method, in particular to establishing a method for protoplast multi-parent fusion based on genome shuffling technology. technical background [0002] Cadmium (Cd) is a heavy metal element with extremely strong biological toxicity. Together with Pb, Cr, As and Hg, it is called the "five poisons" and ranks second. In recent years, the development of modern industry and the wide application of cadmium in smelting, chemical, paper and electronic industries have made Cd pollution more and more serious. Therefore, how to repair cadmium pollution in the environment has become a hot research topic at home and abroad. At present, the research on microbial remediation of cadmium pollution mainly focuses on the selection strategy of cadmium-tolerant microorganisms. [0003] In 2002, Cardayré et al. of Maxgen Company in California, USA proposed the concept of Genome Shuffling technology (translated into "genome shuff...

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Application Information

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IPC IPC(8): C12N15/01C12N15/03C12N15/04C12Q1/04C12R1/74C12R1/125
Inventor 王婷姜春晓孙红文
Owner NANKAI UNIV
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