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Ethanol-tolerant related gene slr0982 of synechocystis 6803 and applications of gene

A technology of slr0982 and Synechocystis, which is applied in the directions of genetic engineering, plant genetic improvement, application, etc., can solve the problems of low bioethanol yield and low ethanol tolerance, and achieves the effect of improving ethanol tolerance and having wide application prospects.

Inactive Publication Date: 2015-01-21
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, the production of bioethanol is relatively small, and there is still a certain gap in industrial large-scale production. One of the limiting factors is the low ethanol tolerance of strains

Method used

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  • Ethanol-tolerant related gene slr0982 of synechocystis 6803 and applications of gene
  • Ethanol-tolerant related gene slr0982 of synechocystis 6803 and applications of gene
  • Ethanol-tolerant related gene slr0982 of synechocystis 6803 and applications of gene

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Construction of Synechocystis 6803 mutant strain △slr0982:

[0017] (1) In vitro amplification of the target gene:

[0018] Gene knockout is achieved by homologous double crossover, and a chloramphenicol resistance gene is introduced at the same time as the knockout gene, and the resistance gene is flanked by upstream and downstream fragments of the gene to be knocked out. The Synechocystis 6803 genome was extracted using a bacterial genome extraction kit as a template, and SEQ ID No.1 and SEQ ID No.2 are respectively the upstream and downstream primers of the gene △slr0982 to be knocked out; SEQ ID No.3 and SEQ ID No.4 is the upstream and downstream primers of the downstream of the gene △slr0982 to be knocked out respectively; PCR amplification is carried out to obtain the upstream fragment and the downstream fragment of the gene △slr0982 to be knocked out; with SEQ ID No.1, SEQ ID No.4 The sequences shown are the upstream and downstream primers, and fusion PCR is per...

Embodiment 2

[0027] The growth curves of wild strain WT and mutant strains in normal liquid medium BG11 and liquid medium BG11 containing ethanol 1.5% (v / v) were determined.

[0028] Synechocystis 6803 (wild strain WT) and the Synechocystis 6803 mutant strain Δslr0982 obtained in Example 1 were cultivated in normal liquid medium BG11 and liquid medium BG11 containing ethanol 1.5% (v / v), and the shaker was set The parameters are light intensity of 2000Lux, rotation speed of 130rpm, and temperature of 30°C.

[0029] The steps are: take OD at the time of inoculation 630nm Add 5 mL of 0.2 fresh cells into 20 mL medium, make 3 parallel samples for each group, measure the absorbance value at a wavelength of 630 nm with a UV-1750 spectrophotometer, measure once every 12 hours, and draw the growth curve ( figure 1 ).

[0030] from figure 1 It can be observed that in the normal liquid medium BG11, the growth status of the wild strain and the mutant strain is almost the same, but in the medium cont...

Embodiment 3

[0032] The growth curves of wild strain WT and mutant strains in normal liquid medium BG11 and liquid medium BG11 containing ethanol 1.8% (v / v) were determined.

[0033] Synechocystis 6803 (wild strain WT) and the Synechocystis 6803 mutant strain Δslr0982 obtained in Example 1 were cultivated in normal liquid medium BG11 and liquid medium BG11 containing ethanol 1.8% (v / v), and the shaker was set The parameters are light intensity of 2000Lux, rotation speed of 130rpm, and temperature of 30°C.

[0034] The steps are: take OD at the time of inoculation 630nm Add 5 mL of 0.2 fresh cells into 20 mL medium, make 3 parallel samples for each group, measure the absorbance value at a wavelength of 630 nm with a UV-1750 spectrophotometer, measure once every 12 hours, and draw the growth curve ( figure 2 ).

[0035] from figure 2 It can be observed that in the normal liquid medium BG11, the growth status of the wild strain and the mutant strain is almost the same, but in the medium ...

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Abstract

The invention discloses an ethanol-tolerant related gene slr0982 of synechocystis 6803and applications of the gene. The gene is shown in SEQ ID No.5. The gene is applied to construction of engineering bacteria for fermenting to produce ethanol. The ethanol-tolerant related gene is identified through constructing mutant strain slr0982 of synechocystis 6803, which has important theoretical and practical significance for improvement of microorganism ethanol tolerance and plays an important role in the fermentation of microorganism; the gene has a wide application prospect.

Description

technical field [0001] The invention belongs to the field of industrial microorganisms, and in particular relates to a synechocystis 6803 ethanol tolerance-related gene slr0982 and its use. Background technique [0002] With the depletion of traditional energy sources, the seriousness of the greenhouse effect, and the continuous deterioration of the environment, bio-energy has attracted more and more attention because of its environmental protection and sustainability. The development and utilization of alternative new energy sources is imminent. As a renewable energy, bioethanol has the characteristics of low pollution, easy transportation and storage, and can compete with gasoline in price. It is the new energy most likely to replace petroleum, and at the same time, it can alleviate the energy crisis to a certain extent. Has great development prospects. [0003] At present, the production of bioethanol is still relatively small, and there is still a certain gap in industr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/31C12P7/06C12R1/01
CPCY02E50/10
Inventor 张亚男陈磊张卫文
Owner TIANJIN UNIV
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