Mutation in plasmid - (Jul/04/2007 )
I have to remove one enzyme site in the plasmid part of the clone. At the moment i donot know this site is present in which part of plasmid, i mean its in ampiciline resistance gene or some origin of replication. I want to ask you that either I have to worried about it can effect transcription processes, i have to remove KPN1 site from the plasmid through one bas pair change, its necessary to bring it back to its origianl or this mutaion
can be harmful during process of stabilizing clone in E.coli for plasmid amplification.
if you delete enzyme site from MCS of vector it would not be a problem, but if the site is in ori or ampr, i also want to know what will happen
check how many KpnI sites are there and where?
As T.reesei wrote, MCS is not a problem. Any other region, i would be cautious, like promoter, intronic sequence, or ampicllin etc.