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Microalgae constitutive expression promoter and application thereof

A constitutive expression and promoter technology, applied in the field of genetic engineering, can solve the problems of limited oil production effect of oleaginous algae and obstacles to the development of microalgae genetic engineering

Inactive Publication Date: 2016-08-10
JINAN UNIVERSITY
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Problems solved by technology

However, the oleaginous effect of oleaginous algae obtained by this method is limited
Currently, in the study of oil production in Phaeodactylum tricornutum, there is only one widely used promoter, which greatly hinders the development of microalgae genetic engineering

Method used

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  • Microalgae constitutive expression promoter and application thereof
  • Microalgae constitutive expression promoter and application thereof
  • Microalgae constitutive expression promoter and application thereof

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Experimental program
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Effect test

Embodiment 1

[0037] 1. Cloning of promoter and GUS gene

[0038] First, the genomic DNA of Phaeodactylum tricornutum was extracted using the plant DNA extraction kit from Magene. The primers were synthesized by Shanghai Sangon Bioengineering Co., Ltd., purified by PAGE method, and diluted to 100 μM when used. The promoter PPtHsp20 was cloned in the DNA of Phaeodactylum tricornutum, and the GUS gene was cloned in the universal vector pCAMBIA1301 (purchased from Shanghai Jielan Biotechnology Co., Ltd.). Wherein the cloning conditions are all adopted PCR method, using KOD-plus-Neo enzyme (TOYOBO, JAPAN) PCR to obtain, the PCR primers of promoter PPt49202 and GUS gene are shown in Table 1, and the PCR reaction system is shown in Table 2 and Table 3 respectively , the promoter of the Pt49202 protein gene of Phaeodactylum tricornutum has not been confirmed. The present invention predicts the core element of the PPt49202 promoter through multiple prediction software, and takes 715bp of it as the...

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Abstract

The invention discloses a microalgae constitutive expression promoter and application thereof .The promoter PPt49202 is cloned out of phaeodactylum tricornutum bohlin, and the nucleotide sequence is shown as SEQ ID NO.1 .The promoter PPt49202 can promote efficient expression, in the growth cycle of algae cells, of downstream genes, and therefore the microalgae constitutive expression promoter can be applied to genetic engineering and has good application prospects in genetic engineering .

Description

Technical field: [0001] The invention belongs to the field of genetic engineering, and in particular relates to a microalgae constitutive expression promoter and its application. Background technique: [0002] At present, the world's energy shortage and environmental pollution have become a huge challenge faced by human society. Therefore, it has become the consensus of all countries in the world to find a new type of renewable and clean energy that can replace fossil energy. Biodiesel, whose performance is basically equivalent to petrochemical diesel, and has good environmental protection and biodegradability, has become the most widely used and fastest growing ideal renewable energy in the world. At present, biodiesel refers to the formation of mono-alkyl fatty acid esters through transesterification of oils (mainly triglycerides) from living organisms and alcohols (methanol or ethanol). [0003] The principle of microalgae to produce biodiesel is to use the photosynthesi...

Claims

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Application Information

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IPC IPC(8): C12N15/113C12N15/79C12N1/13C12R1/89
CPCC07K14/405
Inventor 李宏业刘婉君陈嘉雯杨维东刘洁生
Owner JINAN UNIVERSITY
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