Escherichia coli has been used in genetic engineering for decades, however, its usefulness in the production of bio-based and animal proteins is limited. 2 similar species of earthworms, Eisenia fetida Waki and Eisenia andrei Sagami were used to act as hosts for gene transfers. Both earthworms can be easily obtained and bred, exhibit the production of digestive enzymes and have been noted for being edible- this allows for the possibility of humans ingesting the worms with the recombinant proteins as part of their medicinal plan.
The earthworms were purchased, along with a variety of plasmids. The earthworms were divided into groups of 8 and incubated for 24 hours without food. After this period, the tails were amputated and inoculated on argar plates containing ampicillin ( an antibiotic used to treat several infections). The plates were incubated, after which they were placed in ethanol for 10 minutes. Using a microsyringe, the plasmids (of which one discussed later encoded the luciferase reporter gene luc2) were injected into the amputated tails. Electroporation, a technique where an electrical field is applied to the cell membrane to increase its permeability, was carried out. This procedure allowed the introduction of the DNA into the cell. The fragments were washed, beaten and centrifuged and the supernatant was then collected. A luciferase assay was added to the sample and the luminescence effect (the wavelength-weighted power that a light source emits in a specific direction) was detected using the GloMax-Multi Detection System. The transfection efficiency was determined by dividing the number of samples with positive luciferase activity by the total number of gene-injected samples. A similar experiment was carried out for the other plasmids.
The method developed here is a new host system for the analysis of genes in higher organisms. It uses a transient expression system. While the protein expression rate was not high, the researchers attempted to successfully construct a stable expression system using eggs and earthworm-predictable promoter regions. This work could lead to the development of novel heterogeneous expression systems. While there are many issues associated with the development of transgenic animals, earthworms are relatively low-cost and can be used to feed on waste vegetables. No ethical regulations apply to the use of earthworms. This makes them an attractive target for researchers. The full article can be viewed in the link below.
The earthworms were purchased, along with a variety of plasmids. The eathworms were divided in groups of 8 and incubated for 24 hours without food. After this period, the tails were amputed and inoculated on argar plates containing ampicillin ( an antibiotic used to treat several infections). The plates were incubated, after which they were placed in ethanol for 10 minutes. Using a microsyringe, the plasmids (of which one dicussed later encoded the luciferase reporter gene luc2) were injected into the amputed tails. Electroporation, a technique where an electrical field is applied to the cell membrane to increase its permeability, was carried out. This procedure allowed the introduction of the DNA into the cell. The fragments were washed, beaten and centrifugied and the supernatant was then collected. A luciferase assay was added to the sample and the luminesince effect (the wavelength-weighted power that a light source emits in a specific direction) was detected using the GloMax-Multi Detection System. The transfection efficiency was determined by dividing the number of samples with positive luciferase activity by the total number of gene-injected samples. A simmilar experiment was carried out for the other plasminds.
The method developed here is a new host system for the analysis of genes in higher organisms. It uses a transient expression system. While the protein expression rate was not high, the researchers ateempted to successfully construct a stable expression system using eggs and earthworm-predictable promoter regions. This work could lead to the development of novel heterogeneous expression systems. While there are many issues associated with the development of transgenic animals, earthworms are relatively low-cost and can be used to feed on waste vegetables. No ethical regulations apply to the use of earthworms. This makes them an attractive target for researchers. The full article can be viewed in the link below.
Article:
Akazawa, Si., Machida, Y., Takeuchi, A. et al. Development of a novel heterologous gene expression system using earthworms. Sci Rep11, 8190 (2021). https://doi.org/10.1038/s41598-021-87641-w
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