FRT Stock Info

Kathy Matthews matthewk at fly.bio.indiana.edu
Wed Jul 14 17:58:28 EST 1993


 
Tian Xu has provided the following information for users of his 
FRT stocks.  
 
July 14, 1993 
      
     We have some feedback about the FLP/FRT - Marker strains and 
so we would like to bring three things to the attention of the 
people who are using these strains. Firstly, people in several 
labs have told us that the anti-Myc antibody (c-myc, ab-1) from 
Oncogene Science, Inc. does not work in staining experiments 
involving MYC marker. We have found that indeed this purified 
antibody is now working very poorly in staining MYC marker- 
expressing tissues. We do not know why this is so. Although, we 
did find that this antibody is unstable under some conditions 
after we tried to purify it from tissue culture supernatant. The 
good news is that ascites and supernatants produced from the 
original cell line (Myc 1-9E10.2) give nice staining results for 
many people. People could obtain this cell line from American 
Type Culture Collection (12301 Parklawn Drive, Rockville, 
Maryland 20852) without charge.     
     People should also keep an eye on the FRT stocks that also 
carry the FLP recombinase gene. Since both the FLP recombinase 
gene and its target, the FRT sequences, are present in the same 
flies, it could have a deteriorative effect. For example, these 
stocks may accumulate modifiers which interfere with the function 
of this recombination system. In one case, people have found that 
they are no longer able to induce clones using one of the FLP/FRT 
- Marker strains which was working beautifully when they first 
received it. Since the FLP recombinase gene is under the control 
of the hsp 70 promoter, one way to keep these strains out of 
trouble is to maintain them at 18C. In any case, the strains that 
carry both FRT sequences and the FLP recombinase gene were 
constructed for the connivance of short-term usage. In a long 
term, these stocks can be easily regenerated by crossing the FLP 
gene-carrying chromosomes into the stocks that carry the FRT - 
Marker chromosomes.       
     Please also note a typographical error in the Table 1 of our 
Developmental paper (Development 117, 1223-1237. 1993). The 
abbreviations for the two stocks that carry FRT - NM on the right 
arm of the third chromosome (w; P[ry+; hs-neo; FRT]82B, P[mini- 
w+; hs-NM]88C and w hsFLP1; P[ry+; hs-neo; FRT]82B, P[mini-w+; 
hs-NM]88C) should be 82-NM and 82-NM F instead of 80-NM and 80-NM 
F.  --  Tian Xu


--------------------------------------------------------------
Kathy Matthews			MATTHEWK at FLY.BIO.INDIANA.EDU
Department of Biology		MATTHEWK at INDIANA.EDU
Indiana University              MATTHEWK at IUBACS    
Bloomington, IN  47405-6801     812-855-5782



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