Cloning of flr-1 gene and cDNA.
(A) Genetic and physical map around flr-1
gene. A genomic DNA fragment flanking the transposon Tc1 that caused
the flr-1(ut11) mutation, hybridized to the yeast
artificial chromosome Y50B10 and the cosmid C15A2. An overlapping
cosmid F02D10 and its subclone pMT21–3 rescued the
flr-1 phenotypes by microinjection. The arrow shows the
direction of transcription of the only complete ORF contained in
pMT21–3. Sl, SalI site; Sc, SacI site.
(B, Top) The structure of the 8.5-kb genomic clone
(pMT21–3) that rescued the flr-1 phenotypes. The 17
exons are indicated by boxes. The trans-splice leader SL1 and the
poly(A) tail also are shown. (Middle) The structure of
pMTG25–1, which codes for a GFP(F64L S65T)-tagged FLR-1 protein.
(Bottom) The structure of pMTG24–5, a
flr-1∷GFP(S65C) fusion gene containing a nuclear
localization signal. (C) Rescue of various phenotypes of
flr-1(ut11) by pMTG25–1.