Recombination doubt - Size of a homologous fragment for recombination (Apr/19/2007 )

What is the minimum size of a homologous sequence to be able of recombination in BJ cells (recombination proficient strain)?

I’m constructing shutter vectors to insert different genes of interest and then, recombine with the complete genomes of the viruses (to get the virus with the gene inserted).
I always use 1.2-1.5 kb of homologous sequence per each arm and works perfectly well.
This time, I just have 400 pb , no more (long story).

Do you think 400 pb for one of the arms would be enough for recombination, the other arm is 1 kb??? I’m afraid it could not work, but I don’t know !!

What do you think???

Thanks!!!

Hm,

I've seen successful recombination with arms as short as 50bp, but the efficiency goes down the drain - the guy screened >2000 (sic!) clones to find a good one - but couldn't use a longer arm (long story involved here also...)....

So I'd say this could and should work, but be prepared for a lot of work! - But this should be always the case, anyhow

Mike

Hm,

I've seen successful recombination with arms as short as 50bp, but the efficiency goes down the drain - the guy screened >2000 (sic!) clones to find a good one - but couldn't use a longer arm (long story involved here also...)....

So I'd say this could and should work, but be prepared for a lot of work! - But this should be always the case, anyhow

Mike