Dear colleagues, I work with a bacterium of the genus Helicobacter, and I need to transfect this bacterium with a plasmid. However, few people work with this species, and there is not a single publication that describes the transfection of this bacterium. Therefore, I am trying to find suitable plasmids from Helicobacter pylori. So far, my experiments have been unsuccessful, and I am trying to understand how a plasmid is selected for the bacterium? Ideally, it would be good to predict by bioinformatics, based on the bacterial genome sequencing and the plasmid sequencing. Can anyone suggest what to look out for?
Plasmids can be matched to a particular bacterium by performing a plasmid transformation. This involves introducing the plasmid into a bacterial population and then screening for bacteria that have taken up the plasmid. This can be done using a variety of techniques, such as antibiotic resistance, PCR, or fluorescent markers.
You just need to use compatible vector for the bug.
And I guess you are not correct is saying ".......and there is not a single publication that describes the transfection of this bacterium."
There are many publication related to the transformation of the bug...and these are just top three random hits in the search
Article Helicobacter pylori HP0377, a member of the Dsb family, is a...
https://doi.org/10.1128%2FAEM.01084-07
https://doi.org/10.1128%2FJB.184.10.2755-2766.2002
Try using plasmids with broad host range origins. Else make a synthetic origin of replication based off a filamentous phage origin. To do this you just need to alter the SSO to use the RNA pol III consensus sequence of your target organisms and put the replicase (GPII) under a native host promoter.
Bandla Ramesh
Abhijeet Singh
Dear colleagues, thank you for your answers! I think it is worth clarifying: I work with Helicobacter cinaedi. There is really not a single publication on the transformation of this bacterium. There are only mentions of plasmids found in the H. cinaedi genome sequencing, but these plasmids have not been studied separately, as far as I understand.
I decided to try H. pylori plasmids, since it is a closely related species. However, so far I have the problem that H. cinaedi does not accept H. pylori plasmids , and I am trying to figure out how I can modify the plasmids.
Hence the question of how one can match plasmids to a bacterial species that no one has transfected yet.
I'm hoping that maybe there are methods to predict in silico.
Article Mutations in Genes Encoding Penicillin-Binding Proteins and ...
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