# Protocol for inserting GFP into Lactobacillus strains

**URL:** <https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124>\
**Category:** Kitchen\
**Created:** [April 24, 2025, 11:31am UTC](https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124 "2025-04-24T11:31:04Z")\
**Posts on this page:** 4\
**Page:** 1

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**Author:** ![mamaya](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.hackteria.org/mamaya/32/1025_2.png) [@mamaya](https://forum.hackteria.org/u/mamaya)\
**Post date:** [April 24, 2025, 11:31am UTC](https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124/1 "2025-04-24T11:31:04Z")

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Hello, I am looking for a protocol to insert a GFP plasmid into lactobacillus in agar medium, does anyone has experience in this?

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**Author:** ![Meow](https://avatars.discourse-cdn.com/v4/letter/m/c57346/32.png) [@Meow](https://forum.hackteria.org/u/Meow)\
**Post date:** [April 24, 2025, 11:50am UTC](https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124/2 "2025-04-24T11:50:42Z")

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Do you know what strain you have? A lot of lactobacilli are naturally competent, but not dairy onwe

> **[Unleashing Natural Competence in Lactococcus lactis by Induction of the...](https://pmc.ncbi.nlm.nih.gov/articles/PMC5626998/)**
>
> In biotechnological workhorses like Streptococcus thermophilus and Bacillus subtilis, natural competence can be induced, which facilitates genetic manipulation of these microbes. However, in strains of the important dairy starter Lactococcus lactis,...

Otherwise, your options are going to be doing electroporation

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**Author:** ![mamaya](https://yyz2.discourse-cdn.com/flex030/user_avatar/forum.hackteria.org/mamaya/32/1025_2.png) [@mamaya](https://forum.hackteria.org/u/mamaya)\
**Post date:** [April 24, 2025, 1:44pm UTC](https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124/3 "2025-04-24T13:44:16Z")

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We are trying wild strains from a kimchi batch to see if we can cultivate it and use electroporation to insert a plasmid ring and bring it back to the fermented kimchi. I found this protocol, could this be useful? [https://journals.asm.org/doi/10.1128/aem.02006-10#T2](https://journals.asm.org/doi/10.1128/aem.02006-10#T2)

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**Author:** ![Meow](https://avatars.discourse-cdn.com/v4/letter/m/c57346/32.png) [@Meow](https://forum.hackteria.org/u/Meow)\
**Post date:** [April 24, 2025, 2:05pm UTC](https://forum.hackteria.org/t/protocol-for-inserting-gfp-into-lactobacillus-strains/2124/4 "2025-04-24T14:05:32Z")

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What equipment do you have for electroporation?

I would first start by isolating the bacteria on an agar plate, and when you have isolates, try transforming them, and growing them on selective media. Then, once you get them, re-introduce them into the ferment.

The only problem with using GFP on a plasmid, is that it will lose the plasmid very quickly.