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0343922150
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note
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Caenorhabditis elegans hermaphrodites were observed using a Zeiss Axioplan microscope equipped with a Plan 100 objective and Nomarski differential interference contrast optics. Images were captured with a Hamamatsu Photonics C5810 Color Chilled 3-CCD Camera connected to a Macintosh G3 computer.
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0342616315
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The mig-17 gene was mapped to the right of the Bergerac Tc1 polymorphism bP1 on linkage group V by polymerase chain reaction-sequence tagged site (PCR-STS) mapping. Three-factor mapping placed mig-17 between bP1 and daf-11. Two cosmid clones, T26H10 and F57B7, were found to rescue mig-17(k174). Four coding regions were predicted to lie within the region common to T26H10 and F57B7 by the C. elegans genome sequencing consortium computer analysis. The 5′ terminus of the mig-17 message was analysed using the 5′ RACE System (rapid amplification of cDNA ends; Life Technologies).
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0030589505
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T. G. Wolfsberg and J. M. White, Dev. Biol. 180, 389 (1996); P. Primakoff and D. G. Myles, Trends Genet. 16, 83 (2000).
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Myles, D.G.2
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9
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0343922148
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Mutants were isolated by ethylmethane sulfonate mutagenesis. The mig-17 alleles k135, k147, k167, and k176 were obtained by visual screening of mutants with ventral white-patch phenotypes.
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0343050518
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note
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Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr. X indicates any residue.
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11
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0028365763
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Crabbe, T.1
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Auld, D.S.2
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0028818537
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note
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To construct the mig-17::GFP fusion, a 3645-base pair (bp) Sma I-Afl III mig-17 fragment from the cosmid F57B7 was fused to a Bam HI-Spe I DNA fragment of the GFP plus unc-54 3′ untranslated region from pPD95.75 using appropriate synthetic oligonucleotides. This construct contained 1141 bp of the mig-17 upstream sequence and the 2533-bp mig-77 coding region and all introns. For constructions of unc-54p::mig-17::GFP and lag-2p::mig-17::GFP, the mig-17::GFP coding sequence was placed downstream of the unc-54 promoter from pPD52.102 and the lag-2 promoter from pRB1, respectively. Cosmid clones were co-injected at 10 to 50 μg/ml together with the pMK107 plasmid (100 μg/ml), which contains the EF1α promoter fused to GFP. Plasmid constructs were injected at 100 μg/ml with 50 μg/ml pBR322 either with or without 50 μg/ml unc-119(+) plasmid, pDP#MM016B [M. Maduro and D. Pilgrim, Genetics 141, 977 (1995)].
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0343486358
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Sonicated worm lysates were incubated with anti-GFP monoclonal antibody (Clontech) in a buffer containing 50 mM tris-HCl (pH 8.0), 100 mM NaCl, 10% glycerol, 1% Triton X-100, 5 mM phenylmethylsulfonyl fluoride, 1 μg/ml leupeptin, and 1 μg/ml pepstatin at 4°C for 3 hours. Samples were precipitated with protein A-Sepharose beads (Pharmacia). Immunoprecipitates were subjected to immunoblotting with anti-GFP antibody.
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0343486357
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In mig-17(k174); gon-1(q518) double mutants, DTCs did not migrate at all. In mig-17(k174)/mig-17(k174); gon-1(q518)/+ animals, DTC migration defects of mig-17(k174) mutants were not enhanced.
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0342616314
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We thank A. Coulson, A. Fire, J. Kimble, Y. Kohara, T. Stiernagle, and the Caenorhabditis Genetics Center for materials, and A. Antebi, E. Kipreos, M. Lamphier, and I. Mori for critical reading of the manuscript. Supported by special grants from PRESTO to K.N., and CREST, Uehara Memorial, and Advanced Research on Cancer from the Ministry of Education, Culture and Science of Japan to K.M.
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