transducing recA w/o tetR ?

[R.G. Walters]

In article <CsH3DG.76H at usenet.ucs.indiana.edu> jgraham at bronze.ucs.indiana.edu (the End) writes:
>Has anyone used P1vir to transduce E. coli to recA- using a marker other than
>tn10(tetR) ? My host strain is already tetR and I have been trying to move 
>a tn9-200 linked recA marker from DB1319 (recA938::Tn9-200). So far no 
>luck. 

Many years ago, I did this using a double mutant, recA56 (null) and some
linked marker (nal-resistant? - I forget).  Transduce with selection for
the linked marker, then screen a dozen or so transductants for eg UV 
resistance.

>Does anyone see any inherent problems in this strategy (eg. transducing a 
>tn9 linked mutation) ?

If Tn9 is at all likely to jump after/during transduction, the answer to
your question is yes.  You will need to screen your transductant - you
should anyway, of course :-) - and you may get multiple copies of Tn9
in your new strain.


Robin Walters.                      Robert Hill Institute, Sheffield UK.

A fact is an opinion that everyone agrees with.