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Screening method and application of rapid growth mutants of microalgae

A technology of mutants and microalgae cells, applied in the biological field, can solve problems such as unstable genetics, achieve the effect of solving unstable genetics and avoiding blindness

Active Publication Date: 2017-09-05
GUOTOU BIO TECH INVESTMENT CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0010] In the present invention, the methods of physical and chemical mutagenesis and natural algae screening are combined with population DNA genetic map technology, and by analyzing the genetic background differences between generations, the blindness of continuous generation in conventional algae screening is avoided, and at the same time, the physical The problem of unstable inheritance of chemical mutagenesis mutants

Method used

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  • Screening method and application of rapid growth mutants of microalgae
  • Screening method and application of rapid growth mutants of microalgae
  • Screening method and application of rapid growth mutants of microalgae

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1, activation and pre-cultivation of Haematococcus:

[0034] Pick the Haematococcus NIES-144 monoclonal algae strain from the solid plate and inoculate it in a 50ml Erlenmeyer flask containing 10ml Basal liquid medium, 15μmol·m -2 ·s -1 , and cultured statically at 20°C for 10 days, then transferred to a 250 mL Erlenmeyer flask containing 100 mL of Basal medium and cultured for 4 days under the same conditions. Collect the algae cells by centrifugation at 1500 g for 5 min.

Embodiment 2

[0035] Embodiment 2, EMS chemical mutagenesis algae cell:

[0036] 1. Collect Haematococcus cells in the logarithmic phase by centrifugation at 1500 g for 5 min, and treat with 4% EMS (ethyl methanesulfonate) for 1 h; centrifuge at 1500 g for 5 min to remove the mutagen.

[0037] 2. 50ml sodium thiosulfate Na 2 S 2 o 3 Resuspend the cells to terminate the mutagenesis reaction.

[0038] 3. Centrifuge to remove sodium thiosulfate, wash algae cells twice with Basal medium, and finally resuspend in Basal medium and keep away from light for 24 hours.

[0039] 4. Inoculate the mutagenized algae cells in a 250mL Erlenmeyer flask, and record it as the F0 generation, 15μmol·m -2 ·s -1 , static culture at 20°C, after the F0 generation recovers its vitality and grows fresh algae cells, it is successively subcultured and inoculated, which are respectively recorded as F1, F2, F3....

Embodiment 3

[0040] Embodiment 3, inferring the appropriate number of passages by calculating

[0041] Assume an initial inoculum of N cells containing a fast growing mutant.

[0042] The wild type can grow 10 times in the logarithmic phase, assuming that the fast growing mutant can grow 20 times in the logarithmic phase, then the number of passages required when the proportion of the fast growing mutant increases to 50% can be calculated.

[0043] The proportion of fast-growing mutants (Fn) when each generation grows to the logarithmic phase is as follows

[0044]

[0045] When Fn=50%, if the initial inoculation amount N=4X10 4 , then it can be deduced that the number of passages n=13.7. However, due to the uncertainty of the growth efficiency of the mutants obtained after mutagenesis and the possible differences in the initial inoculum volume, there will be large differences in the number of passages required in practice and in theoretical calculations. There is therefore still a n...

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Abstract

The invention specifically relates to a screening method and application of microalgae mutant strains of high growth rates, especially to a method for screening rapid growth mutants of microalgae via a combination of population DNA genetic mapping technology AFLP and flow cytometry, belonging to the field of biotechnology. The method provided by the invention is rapid in screening and reliable in results.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a screening method and application of microalgae mutant strains with high growth rate. In particular, it relates to a method for screening microalgae fast-growing mutants by combining population DNA genetic mapping technique AFLP and flow cytometry. The method is rapid in screening and reliable in results. Background technique [0002] Microalgae belong to ancient lower plants and are widely distributed in lakes, oceans and other waters. They have the ability to efficiently absorb solar energy and fix carbon dioxide for rapid proliferation (photosynthesis). At the same time, microalgal cells can produce a variety of high-value nutrients and chemical raw materials, such as carotenoids, proteins, polysaccharides, oils, etc. With the rapid development of the economy, the problem of energy shortage needs to be solved urgently. After two generations of biomass energy develo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/01C12R1/89
CPCC12N15/01
Inventor 韩丹翔宋晓瑜胡强
Owner GUOTOU BIO TECH INVESTMENT CO LTD
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