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Application of large-sized chlorophta chlorophyll body as exogenous gene receptor and expression vector

A technology of exogenous gene and expression vector, which is applied in the application field of receptors and expression vectors, can solve the problems of lack of performance and no start of macroalgae chloroplast genetic transformation system, and achieve low cost, large amount of exogenous gene products, and multiplication The effect of high ability

Inactive Publication Date: 2009-05-06
YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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Problems solved by technology

However, there are very few related studies in our country, and the establishment of a chloroplast genetic transformation system for macroalgae has not yet begun. As the marine algae with the largest biomass on the earth, it is far from playing its due role in the application of plant genetic transformation.

Method used

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  • Application of large-sized chlorophta chlorophyll body as exogenous gene receptor and expression vector
  • Application of large-sized chlorophta chlorophyll body as exogenous gene receptor and expression vector
  • Application of large-sized chlorophta chlorophyll body as exogenous gene receptor and expression vector

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Embodiment

[0022] experiment method:

[0023] The polyethylene glycol (Polyethylene glycol, PEG) method in the protoplast aggregation and regeneration process of Paleopsis algae;

[0024] Plasmid construction:

[0025] BH 7 As a basic construction, construct a plasmid BH with only an enhanced fluorescent reporter gene 7 P (EGFP is the target gene, namely enhanced green fluorescent protein gene). Plasmid DNA extraction and purification, restriction endonuclease reaction, DNA fragment recovery, T4 DNA ligase reaction, plasmid transformation, and screening and identification of recombinants were all carried out in accordance with the "Molecular Cloning Experiment Guide".

[0026] The vector used for site-directed transformation of the chloroplasts of Paleopsis chloroplast uses two genes rps7 (ribosomal protein small subunit gene 7) and ndhB (NADH dehydrogenase subunit B gene) as homologous recombination fragments. The EGFP gene is inserted between the homologous recombination fragments,...

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Abstract

The invention relates to a chloroplast genome of chlorella leaves, in particular to application of chloroplast of large chlorella leaves as a receptor and an expression vector of exogenous genes. The application has the following advantages: a conversion system and a conversion method are integrated, processes are simple and convenient, conversion efficiency is high, and culture conditions are simple; the exogenous genes have wide receiving range and large yield, and can be expressed in a shorter time; and the cost is low, and the application effect is good.

Description

technical field [0001] The invention relates to a green algae chloroplast genome, in particular to the application of a large green algae chloroplast as an exogenous gene receptor and expression carrier. Background technique [0002] With its unique advantages, plant genetic engineering has become one of the important ways to solve global food problems, environmental crisis, medical and health problems. As a new genetic transformation system, chloroplast has received more and more attention, and chloroplast transgenic plants through genetic engineering have become a new type of bioreactor. [0003] Chloroplast transgenic plants have many advantages as receptors and expression vectors of exogenous genes: 1) As a genetic transformation system, chloroplasts can not only achieve a large amount of stable expression of exogenous genes, but also form correct disulfide bonds and spatial conformation of exogenous proteins. ; 2) The high copy number of the genome of chloroplasts prov...

Claims

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Application Information

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IPC IPC(8): C12N15/82
Inventor 叶乃好庄志猛王清印毛玉泽
Owner YELLOW SEA FISHERIES RES INST CHINESE ACAD OF FISHERIES SCI
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