I would like to transfect bacteria with some plasmids. However, the bacteria are not competent but either isolated from the environment or purchased reference strains.
Can I transfect these strains?
Is there a kit or an established protocol?
Is the procedure much more different or the efficiency much lower than with competent cells?
Thank you
Hi, I don't have any experience with this, but just last week I saw this preprint
https://www.biorxiv.org/content/10.1101/2024.05.27.596136v1
Maybe it could be of interest to you.
Wish you luck. I'd love to hear your progress!
Thank you. It's interesting but a bit too fringe yet. I was looking for something more mainstream (easy to do).
Typically putting plasmids into bacteria is called transformation, not transfection, just FYI to help you find literature easier.
Non-model bacteria are typically transformed either by electroporation at high voltage with high resistance settings, or by conjugation with a donor strain. The exact details depend a lot on what specific type of bacteria you're working with. Bacterial physiology is quite variable between distantly related species so it's hard to generalize a transformation protocol. For example, many Gram-positive bacteria need to have their cell walls weakened in a high osmolarity solution (to prevent lysis) before they can be transformed by electroporation, whereas Gram-negative bacteria do not have this issue. Some bacteria have large polysaccharide capsules that need to be removed or inhibited before they can be efficiently transformed. That's not getting into the fact that all plasmids have host range/expression limitations.
I would start by searching around for transformation protocols of the most closely related species to the ones you're interested that you can find.
From Britannica (former Encyclopedia Britannica) "transfection, technique used to insert foreign nucleic acid (DNA or RNA) into a cell... Transfection is employed in a variety of research fields, including microbiology and genetics" (https://www.britannica.com/science/transfection).
"Transformation, in biology, one of several processes by which genetic material in the form of “naked” deoxyribonucleic acid (DNA) is transferred between microbial cells." (https://www.britannica.com/science/transformation-biology)
The latter typically involves the intake of DNA fragments; the NATURAL intake of plasmids is called conjugation. (https://www.britannica.com/science/conjugation-sexual-process).
PS: I got a protocol from ThermoFisher
Luigi Marongiu maybe try something more science-y next time.
https://pubmed.ncbi.nlm.nih.gov/?term=bacteria+transformation
https://pubmed.ncbi.nlm.nih.gov/?term=bacteria+transfection
transfection is usually used for animal cells, while transformation is used for bacterial (and plant) cells.
I'm curious, what is the name of the protocol from ThermoFisher?
Luigi Marongiu the answer from Alexandria is good especially with regard to electroporation. I would suggest that approach as otherwise you may need to do a lot of optimiazation for making chemically competent cells.
However you need to pay attention to what it is that you are transforming. Most plasmids are not broad host range, although a few are. So if you are planning to transformn a plasmid be sure it will replicate (or integrate).
On a related note as Tomáš Hluska pointed out, the term transfection is commonly used for animal cells and transformation for bacteria. With bacteria the term transfection has historically been used when being transformed by viral DNA and transformation by plasmid DNA. Conjugation is the natural exchance of plasmids (or genomes) by cell to cell contact, however you can have natural transformation as well which is the uptake of free DNA.
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