How to develop drug resistance in bacteria? - (Aug/25/2008 )
Hello, can some one help me in this topic?
I need to develop a resistant variety of bacteria using some antibiotics. How shall i go about this? Plz help me!
You can always use a plasmid with an antibiotic resistance marker gene. Just perform a transformation. For most gram negative bacteria a chemical transformation should be enough and is fairly easy to do. For gram positive bugs probably you will need electroporation.
I am not so sure what you really need and how you are supposed to perform your experiment anyway. We can use a little more of information to help you 
I am not so sure what you really need and how you are supposed to perform your experiment anyway. We can use a little more of information to help you
Thanks for your kind reply.
It is been told by one of my colleague that, growing bacteria in 1/4th MIC in a broth overnight and chk for growth(by plating) and continous incubation for 2 more days, i can get the resistant variety. Is this methodology right?
The antibiotic which is asked to use is Trimethoprim and the organism is a gram negative bacterium in my case, i dont think there is a plasmid with this antibiotic marker. I am totally new to this kindly help mt in this issue. Thanks in advance.
Trimethoprim resistance involves a mutation in the dihydrofolate reductase gene. So, your protocol is trying to enrich for cells containing spontaneous mutations in this gene. The procedure sounds reasonable enough; it is the rate at which spontaneous mutations occur in your culture population in general and the chances of enriching for a spontaneous dihydrofolate reductase mutant that will be limiting.
In general, though, I think the approach is sound.
In general, though, I think the approach is sound.
Thanks man. Will let u know d result guys.