发信人: leohawk (leohawk), 信区: Biology
标 题: Re: question about P element transposition....
发信站: The unknown SPACE (Wed May 9 18:44:35 2001), 转信

I have some experience in this, not so good experiences:

local hopping using P element depends a LOT on the specific P element you
are dealing with, some hop well, some never hop. I spent more than half a year
trying to hop one at one point, never give me deletion in the gene,(only 52
bp away),I have to use EMS to get my mutant later.

another guy was lucky, he hopped several times and got a deletion (in another
gene). So, if you can't make it work after a while, try something different.

One thing you should pay a lot of attention, when you hop the P element away,
look not only at the gene you are interested in, but also the gene on the
other side of the P element insertion, make sure that gene is intact, and its
5' or 3' regulation sequence is not affected.

GOOD LUCK, you may need it.



【 在 gilmer (height) 的大作中提到: 】
: Anyonw familiar with P element transposition?
: I tried to do some local hopping to get a new insertion in a certain gene,
: but I failed in getting new inwertion that showed the mutant phenotype.
: Actually I got some hopping that produced mutant phenotype, I think there
: were some excision of the p element. I tried to map the possible imprecise
: excision around the original p element inersion site, but found nothing.
: The question is: will the deletion caused by p element hopping be far away
: from the original insertion site? ( I suppose the frequency of multiple
: hopping to be very low, but not sure. Actually I got quite a few
: non-reinertion hopping flies. But I am not sure if they were the result of
: multiple hopping).


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