Blue-green alga gene transfer method favorable to harvest

A technology of gene transfer and cyanobacteria, which is applied in genetic engineering, plant genetic improvement, botany equipment and methods, etc., can solve the problems that limit the industrialization of cyanobacteria genetic engineering technology, cannot be collected by filtration, and are not easy to collect algae cells, etc. , to achieve the effects of easy harvesting, preservation and transportation, and low production cost

Inactive Publication Date: 2005-04-20
INST OF AQUATIC LIFE ACAD SINICA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

They grow uniformly and dispersed, which is convenient for counting and quantification, and is suitable for research work, but it is difficult to carry out large-scale production in an open system, and the cells or filaments are small and cannot be collected by filtration, even if they are produced, they are not easy to collect algae cell
These difficulties have become a bottleneck restricting the industrialization of cyanobacteria genetic engineering technology. Therefore, if a gene transfer method can be established in cyanobacteria that can grow in agglomerates and float, it will be a breakthrough in cyanobacteria genetic engineering.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Example 1: Cloning of a fragment of genetic material containing the B. sphaericus binary toxin gene into a shuttle plasmid. According to the overall plan, the shuttle plasmid, driver plasmid and helper plasmid cloned with the binary toxin gene were incorporated into the same Escherichia coli cell, and then conjugatively transferred with Nostoc griseus UTEX1037 to obtain neomycin-resistant algal colonies. The resistant algae colonies were inserted into the BG11 culture medium containing neomycin, and after one month, healthy growth algae agglomerates could be seen floating on the surface of the culture medium. Under the condition of aerated culture, the transgenic Nostoc algae still formed lumps. Therefore, when producing transgenic Nostoc cinereus on a large scale, if static culture is used, it can be directly fished from the surface of the culture tank; if aeration culture is used, nylon nets can be used to collect algae blocks.

[0019] Due to the introduction of the...

Embodiment 2

[0020] Example 2: The strong promoter of the thioredoxin gene device was cloned into pRL25, and introduced into Nostoc griseus according to the general scheme and the gene transfer method described in Example 1. This genetically modified algae is produced and harvested in large quantities for the preparation of crude thioredoxin and as an additive in the treatment of dairy products to improve digestion and prevent allergies.

Embodiment 3

[0021] Example 3: The strong promoter of the synthetic human epidermal growth factor gene device was cloned into the shuttle plasmid pRL25. The gene transfer method described in Example 1 was followed to introduce No. griseus. This transgenic algae is mass-produced and harvested for the preparation of crude epidermal growth factor, which is used as an additive in cosmetics to promote skin metabolism and wound healing.

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PUM

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Abstract

The invention discloses a gene transfer method for cyanobacteria that is beneficial to harvesting, and relates to a gene transfer method for microorganisms, in particular to a gene transfer method for introducing an exogenous gene into a filamentous nitrogen-fixing cyanobacteria capable of agglomerating growth. In order to overcome the non-caking problem of cyanobacteria currently used to express foreign genes, a filamentous nitrogen-fixing cyanobacteria that grows in caking and floating growth was screened from nearly 70 algae strains to establish a gene transfer system, so that the algae can be obtained from Fishing directly from the culture solution or harvesting by filtering with ordinary nylon mesh is beneficial to the industrialization of cyanobacteria genetic engineering. In order to adapt to the gene transfer of this kind of cyanobacteria, important improvements have been made to the combination mode of cyanobacteria.

Description

technical field [0001] The invention relates to a gene transfer method for microorganisms, in particular to a gene transfer method for introducing an exogenous gene into a filamentous nitrogen-fixing cyanobacteria capable of agglomerating growth. technical background [0002] Genetic engineering can be used to express valuable genes in microorganisms, animals and plants to produce genetically encoded products, or to modify the traits of microorganisms, animals and plants. Commonly used microorganisms include Escherichia coli, Bacillus, and yeast. The gene transfer and expression systems of these microorganisms have been matured after years of development, and have been successfully used in the production of insulin, interferon, interleukin and other drugs. It has the advantages of high expression, perfect protein extraction and purification methods, etc.; but the high cost of cultivation can only be applied to the production of expensive drugs. If it is used in the field o...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/13C12N1/21C12N15/12C12N15/63C12N15/70
Inventor 徐旭东孔任秋
Owner INST OF AQUATIC LIFE ACAD SINICA
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