A lot of mutation methods are PCR-based and the mutations in vectors are then transferred to host cells. Are there any ways to mutate a plasmid in the bacteria in vivo? I want to make random mutations in just plasmid to express higher performance protein.
Many classical drug and radiation methods from the 60s to 80s in literature.
May be if u want to introduce random mutations, you can use GeneMorph-II, or by exposing to UV radiation.
and for mutations of interest, u can go by Site directed mutagenesis or transpososn mediated mthod.
You can keep plasmid containing bacterial cells with 1% ethylmethanesulphonate (EMS) for 1 or 2 hrs to generate random mutations in plasmid.
I have used pentapeptide scanning mutagenesis which was effective and I got some mutants through this approach.
For introduction of random mutation in genomic bacterial DNA I used EZ-Tn5™ Tnp Transposome Kit, Epicentre Biotechnology or transduction with P1vir. In case of single nucleotide mutation in plasmid mutagenesis I used Site directed mutagenesis kit and PCR.
YOU CAN USE A CHEMICAL MUTAGEN (EMS, NTG), HOWEVER THE MUTATIONS CAN BE GENERATED IN CHROMOMOSE BESIDES THE PLASMID.
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