Screening method of chlamydomonas reinhardtii selenium-rich mutant

A screening method, the technology of Chlamydomonas reinhardtii, which is applied in the field of screening of selenium-enriched mutants of Chlamydomonas reinhardtii, can solve the problems of lack of screening methods for selenium-enriched mutants of Chlamydomonas reinhardtii, and achieve the effect of high screening efficiency and simple steps

Inactive Publication Date: 2017-03-22
JIANGHAN UNIVERSITY
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Problems solved by technology

[0006] The invention provides a screening method for a selenium-enriched mutant of Chlamydomonas reinhardtii, which solves the technical problem of lack of a screening method for a selenium-e

Method used

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example 1

[0075] 1) Set the concentration of SeO2 as 0.05mmol / L, 0.1mmol / L, 0.2mmol / L, 0.4mmol / L, 0.8mmol / L, 1.0mmol / L, 5.0mmol / L, 10.0mmol / L respectively, with Without SeO 2 For control.

[0076] 2) Inoculate an appropriate amount of Chlamydomonas cells into R liquid medium, culture for 3 days at 20°C-25°C under a photoperiod (14h light / 10h dark), and transfer the Chlamydomonas cells grown to the logarithmic phase to M Carry out synchronous culture in liquid medium for 48-72h;

[0077] 3) Count the synchronized Chlamydomonas cells.

[0078] 4) Take 5ml of counted Chlamydomonas cells into a centrifuge tube, centrifuge at 2500rpm for 3min, remove the supernatant, and resuspend the cells with an appropriate amount of M medium to make the concentration of Chlamydomonas cells reach 1×10 8 cells / ml.

[0079] 5) Take 50 μl of Chlamydomonas cells with the above concentrations, and drop them into 2 Concentration of TAP solid plate, observe the growth of Chlamydomonas.

[0080] After the C...

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Abstract

The invention discloses a screening method of a chlamydomonas reinhardtii selenium-rich mutant. The screening method comprises selecting an appropriate screening concentration of a selenium-rich mutant according to influence produced by different concentrations of SeO2 on the growth of chlamydomonas reinhardtii, acquiring a chlamydomonas reinhardtii mutant, screening a selenium-rich mutant, and identifying a mutant gene of the screened selenium-rich mutant. The screening method has simple processes and high screening efficiency, and realizes fast and accurate screening of a chlamydomonas reinhardtii mutant capable of growing at a high selenium concentration. Through RESDA-PCR, the mutant gene and the inserted mutation site can be identified.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a screening method for a selenium-enriched mutant of Chlamydomonas reinhardtii. Background technique [0002] Selenium is an essential trace element for biology. If the human body is deficient in selenium for a long time and cannot be supplemented, the metabolic process related to selenium will be blocked, which will lead to the occurrence of certain diseases. Trace amounts of selenium can improve human immunity, stimulate the body to produce immunoglobulins and antibodies, and enhance specific immunity. At the same time, selenium also has anti-oxidation effect, which is an important component of antioxidant enzyme glutathione peroxidase, which can remove excessive active oxygen free radicals in the body, cut off peroxidation reaction, and protect the structure of cell membrane and chromosome. It is destroyed, and protects the structure and function of macromolecular bioactive subst...

Claims

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Application Information

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IPC IPC(8): C12N15/01C12Q1/68C12Q1/04
CPCC12N15/01C12Q1/6858
Inventor 李璨彭海胡长峰李丽丽
Owner JIANGHAN UNIVERSITY
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