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Beaton, A. (2000.1.31). BDGP P element Update. 
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FBrf0126832
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Personal communication to FlyBase
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Text of Personal Communication
From abeaton@XXXX Mon Jan 31  20:13:33  2000
To: flybase-updates@XXXX
Dear FlyBase,
Hi. I hope the work on these Ps can be included in your records.
Respectfully submitted,
Amy Beaton
for BDGP
BDGP P element Update
49 P element strains were selected for excision work. Their genomic
flanks associate them with ESTs that have homologies with interesting
transcripts in other organisms.
l(2)00734 57E3-4
l(2)01501 32D1-2
l(2)02516 48C1-2
l(2)03775 46D1-2
l(2)04524 42F1-2
l(2)06444 48E10-11
l(2)07838 55D1-2
l(2)09373 60B10-11
l(2)10408 35B8-9
l(2)k00103 51C1-2
l(2)k00311 31E1-2
l(2)k00619 21F1-2
l(2)k03902 36B1-2
l(2)k04810 53E1-2
l(2)k05103 47F8-9
l(2)k06103 47F1-2
l(2)k06109 42A1-2
l(2)k06709 31D8-9
l(2)k06710 36C8-11
l(2)k07736 23F5-6
l(2)k07918 22B6-7
l(2)k08819 36A12-14
l(2)k09003 25C1-2
l(2)k09217 52D11-12
l(2)k09514 44C1-2
l(2)k10617 27C6-8
l(2)k10712 49D5-6
l(2)k13807 32D4-5
l(3)00848 99F10-11
l(3)01031 94F1-2
l(3)01418 64C9-10
l(3)02267 84C1-2
l(3)04210 89A1-2
l(3)05146 97D3-6
l(3)05614 84A4-5
l(3)10547 73D1-2
l(3)j10B2 77B6-7
l(3)j1C7 66B10-11
l(3)j1E6 82A3-5
l(3)j2B8 67B10-11
l(3)j2D9 96B19-20
l(3)j3A4 82D1-2
l(3)j3B9 100B2-4
l(3)L2249 65E10-11
l(3)L4032 89D1-2
l(3)L4910 94B4-5
l(3)rK561 66D10-13
l(3)rL562 67E1-4
l(3)s1939 72D8-9
5 independent sublines (single chromosomes) of each strain were
outcrossed to either w1118 or ry506 for six generations before
rebalancing crosses were made to reestablish the stocks from single
chromosomes.
The following strains were found to bear viable insertions and should
be changed to no P phenotype.
l(2)k04810 53E1-2 homozygotes (k10209 and k04810 inserted one base apart)
l(2)k05103 47F8-9 homozygotes
l(2)k07736 23F5-6 homozygotes \** see note below regarding background lethals
l(2)k09217 52D11-12 homozygotes
l(3)05146 97D3-6 homozygotes. It seems likely that the
l(3)05146 stock I started with had a problem because there are
multiple alleles of the locus and another one l(3)L1602 has remained
lethal through outcrossing.
One line was not recovered after outcrossing:
l(3)rK561 66D10-13 Rebalancing of outcrossed males via e
ftz ry506/TM3, ryRK resulted in no male, ry+/TM3 progeny in 4 of
5 sublines. The 5th subline gave a mess in the F2.
The remainder of the lines were still lethal after outcrossing.
Excision work proceeded with lines that were either single insert
lines or were verified only by means of noncomplementation with a
deficiency. 50 independent excision events were followed per line
(with the exception of l(3)j1C7 which needed 100 lines to get 6
viable lines). The following lines were successfully reverted and the
frequency of reversion is indicated
l(2)00734 08.6%
l(2)07838 66.7%
l(2)09373 35%
l(2)10408 61.1%
l(2)k00311 77.6%
l(2)k00619 65%
l(2)k06103 74.4%
l(2)k09514 27.7%
l(2)k10712 50%
l(2)k13807 74%
l(3)01031 19.35%
l(3)01418 54.83%
l(3)04210 11.76%
l(3)j10B2 67.85%
l(3)j1C7 06%
l(3)j1E6 15.15%
l(3)j2B8 39.28%
l(3)j2D9 69.23%
l(3)j3A4 80.00%
l(3)L4032 67.56%
l(3)L4910 18.75%
l(3)rL562 44.73%
Rebalancing of excision chromosomes failed to produce homozygous
viable chromosomes in the following lines.
l(2)k07918 no target site duplication
l(3)L2249
l(3)s1939
l(3)02267 no target site duplication
l(3)s1939 had 26 excisions and none produced homozygous viable chromosomes.
l(3)02267 had 29 excisions and none produced homozygous viable chromosomes.
The take home message from this work: Lethal P lines are likely to
be lethal due to the insertion if they are still lethal after
outcrossing for a number of generations. Excision work is relatively
tedious and outcrossing itself does the job.
>From rd120@XXXX Thu Mar 02  16:18:43  2000
>To: abeaton@XXXX
>Subject: Re: your pc
Background lethals and l(2)k07736:
You wrote:
>The following strains were found to bear viable insertions and should
>be changed to no P phenotype.
..
>l(2)k07736 23F5-6 homozygotes
..
so I am inclined to separate the lethality from the insertion i.e.
P{lacW}Phas1k07736 will stay P{lacW}Phas1k07736 but will become
marked as a viable insertion - presumbly the chromosome is still lethal
so I will invent l(2)k07736k07736 with no associated P insertion, But
(AND THIS IS THE PROBLEM) there are two other alleles of Phas1 that are
also lethal, k07730 and k07739, both of these are said to fail to
complement k07736. Is it likely that 3 inserts in Phas1 are all viable
and all have a second hit in 'l(2)k07736'? Are these possibly a
cluster with the same background lethal? The numbers are very close
together, for what it is worth.
From abeaton@XXXX Fri Mar 03  18:38:05  2000
To: Rachel Drysdale (Genetics) <rd120@XXXX>
Subject: Re: your pc
that's exactly right. In l(2)k***** names the three digits following
the k indicate the 'pool' number ie the bottle number of the cross
and these need to be treated as a potential premeiotic cluster. The
final two digits just count which one from the bottle. ... So
yes all three k077* are a cluster with a shared background lethal.
that's it,
Amy
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