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Method for effectively screening out chlamydomonas hydrogen desorption mutant

A mutant, Chlamydomonas technology, applied in the field of screening Chlamydomonas hydrogen-release mutants, can solve problems such as unclear mechanism and regulation method, ineffective genetic transformation, and restrictions on the application of hydrogen production pathways by microalgae, so as to reduce the workload , the effect of improving efficiency

Inactive Publication Date: 2013-05-29
INST OF BOTANY CHINESE ACAD OF SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because photosynthetic water releases oxygen and produces hydrogen are two complex biological metabolic processes that are both contradictory and closely related in microalgal cells, and the mechanism and regulation of these processes are still unclear, resulting in ineffective genetic modification. However, the efficiency of conversion of light energy to hydrogen cannot be significantly improved, which seriously restricts the application of microalgal hydrogen production from the source.

Method used

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  • Method for effectively screening out chlamydomonas hydrogen desorption mutant
  • Method for effectively screening out chlamydomonas hydrogen desorption mutant
  • Method for effectively screening out chlamydomonas hydrogen desorption mutant

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Embodiment 1

[0022] 1. Plant material: Chlamydomonas reinhardtii (Chlamydomonas reinhardtii), algae species CC425 (arginine-deficient, Arg-) from Duke University algae library Chlamydomonas reinhardtii (Chlamy center, http: / / www.chlamy.org / ) were purchased; other mutants were obtained by our laboratory using T-DNA random insertion method.

[0023] 2. Plasmid: pJD67 plasmid was purchased from Duke University Chlamydomonas Center (Chlamy center, http: / / www.chlamy.org / ); The pJD67 plasmid is transformed from the commercialized pBluscript SK plasmid, with a full length of 11284bp, containing the gene of spermylsuccinate lyase and the gene of ampicillin resistance. When transforming algal strains, linearize with KpnI to improve the transformation efficiency. The sequence of pJD67 is sequence 1, and the structure diagram of pJD67 is as follows Figure 4 .

[0024] 3. Medium

[0025] 1) Normal TAP medium: NH 4 C1 0.4g / L; MgSO 4 ·7H 2 O 0.1g / L; CaCl 2 2H 2 O 0.05g / L; K 2 HPO 4 0.108g...

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Abstract

The invention discloses a method for effectively screening out a chlamydomonas hydrogen desorption mutant, which comprises the following steps of: (1) introducing pJD67 plasmid to Chlamydomonas reinhardtii CC425, culturing transformants on a culture medium without arginine, and screening out the transformants which can grow, i.e., the mutant embedded with the pJD67 plasmid; and (2) screening out in the mutant embedded with pJD67plasmid to obtain a transgenic chlamydomonas mutant with increased hydrogen desorption quantity. According to the invention, the workload of directly measuring the hydrogen generation quantity is reduced; the efficiency of screening out the chlamydomonas hydrogen desorption mutant is increased; an important foundation for study on a mechanism of increasing the hydrogen generation quantity by microalgae photosynthesis is established; and meanwhile, the basis of improving a large-scale hydrogen generation technique in the future is provided.

Description

technical field [0001] The invention relates to a method for screening hydrogen-releasing mutants of Chlamydomonas. Background technique [0002] Due to the depletion of mineral resources, looking for clean alternative energy has become an urgent issue. Hydrogen is the simplest and most abundant element in the universe. Its calorific value is as high as 118.4kJ / g, and only water is produced after combustion. It is generally considered to be the most attractive alternative energy source. [0003] However, the concentration of hydrogen on the earth's surface is less than 1mg / L, which only accounts for a very small part of the earth's surface atmosphere. Most of the hydrogen in nature exists in a combined state. The traditional chemical hydrogen production method uses electrolysis of water or pyrolysis of petroleum. , natural gas, production costs are generally higher. Therefore, new hydrogen production methods, especially those that can utilize renewable energy and produce h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/13C12N15/63C12R1/89
Inventor 黄芳赵磊陈梅孙永乐
Owner INST OF BOTANY CHINESE ACAD OF SCI
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