1
$\begingroup$

How do delete a single gene encoded in an operon on a plasmid using E. coli? Could you use the same principle as when knocking out a gene from the bacterial chromosome which is e.g. suicide vector-derived allelic replacement through homologous recombination?

Thanks for your response.

|improve this question
$\endgroup$
  • 1
    $\begingroup$ So you have a plasmid and you want to delete a section? Why not do the deletion outside the cell with PCR or something and then transform into new cells? $\endgroup$ – user137 Jun 6 '15 at 14:40
1
$\begingroup$

Well the simplest way is to cut the gene at the both ends from operon by means of proper restriction enzymes but in case that you get more than one recognition site then you can use microRNA or proper oligo to make a hairpin loop and break that part If you only want to silence the operon then you just need to cut it nearly at middle of the gene and ligate it with a nonsense sequence, then it will loose it's functionality.

|improve this answer
$\endgroup$

Your Answer

By clicking “Post Your Answer”, you agree to our terms of service, privacy policy and cookie policy

Not the answer you're looking for? Browse other questions tagged or ask your own question.