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Volumn 274, Issue 5291, 1996, Pages 1389-1391

Control of C. elegans larval development by neuronal expression of a TGF-β homolog

Author keywords

[No Author keywords available]

Indexed keywords

PHEROMONE; RNA; TRANSFORMING GROWTH FACTOR BETA;

EID: 10544256181     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.274.5291.1389     Document Type: Article
Times cited : (439)

References (31)
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    • W. B. Wood, Ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
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    • Cosmid DE9 [D. Pilgrim, Genome 36, 712 (1993)] transformed a daf-7(e1372) mutant strain to wild type, indicating that it carried daf-7(+). A 10-kb Cla I-rescuing fragment spanned the site of a Tc1 insertion in daf-7(m434::Tc1) as determined by Southern (DNA) blot analysis of mutant and parent daf-7(+) genomic DNA. A 5-kb Cla I-Pst I fragment DR#227 (Fig. 1A) from the 10-kb fragment was microinjected into the germlines of daf-7 mutants e1372 and n696 along with plasmid pRF4 [ominant mutant rol-6(su1006)] as a transformation marker (7). Rescued progeny were identified as L4 and adult rollers at 25°C. Five cDNA clones were identified through use of a 1-kb Bam HI fragment from DR#227 (Fig. 1A) to probe a mixed-stage C. elegans cDNA library [R. J. Barstead and R. H. Waterston, J. Biol. Chem. 264, 10177 (1989)]. A 1.2-kb, full-length cDNA (DR#231) and the entire 5-kb genomic fragment (DR#227) were sequenced.
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    • Cosmid DE9 [D. Pilgrim, Genome 36, 712 (1993)] transformed a daf-7(e1372) mutant strain to wild type, indicating that it carried daf-7(+). A 10-kb Cla I-rescuing fragment spanned the site of a Tc1 insertion in daf-7(m434::Tc1) as determined by Southern (DNA) blot analysis of mutant and parent daf-7(+) genomic DNA. A 5-kb Cla I-Pst I fragment DR#227 (Fig. 1A) from the 10-kb fragment was microinjected into the germlines of daf-7 mutants e1372 and n696 along with plasmid pRF4 [ominant mutant rol-6(su1006)] as a transformation marker (7). Rescued progeny were identified as L4 and adult rollers at 25°C. Five cDNA clones were identified through use of a 1-kb Bam HI fragment from DR#227 (Fig. 1A) to probe a mixed-stage C. elegans cDNA library [R. J. Barstead and R. H. Waterston, J. Biol. Chem. 264, 10177 (1989)]. A 1.2-kb, full-length cDNA (DR#231) and the entire 5-kb genomic fragment (DR#227) were sequenced.
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    • Barstead, R.J.1    Waterston, R.H.2
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    • note
    • Polymerase chain reaction (PCR) was used to amplify the entire daf-7 gene of each mutant. Gel-purified PCR products were subcloned into pBluescript. At least two independent PCR clones per mutant were sequenced.
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    • S. Daopin, K. A. Piez, Y. Ogawa, D. R. Davies, Science 257, 369 (1992); D. L. Griffith et al., Proc. Natl. Acad. Sci. U.S.A. 93, 878 (1996).
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    • Griffith, D.L.1
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    • PCR was used to create an Xma I site immediately before the daf-7 start codon. daf-7p::gfp (DR#271) was constructed by subcloning the 3-kb Xba I-Xma I fragment 5′ of the daf-7 start codon from DR#227 (Fig. 1A) into gfp vector Tu#61 [M. Chalfie, Y. Tu, G. Euskirchen, W. W. Ward, D. C. Prasher, Science 263, 802 (199-4)]. DR# 271 and pRF4 plasmid were coinjected into wild-type animals (8). Lines carrying the transgenes stably integrated into a chromosome were generated by γ-irradiation (76). Neuron identification was based on the position of nuclei and axonal morphology (15). GFP expression was observed in growth-promoting conditions at 15°C, 20°C, and 25°C. Faint GFP expression in the AIM interneurons was observed at low frequency.
    • (1994) Science , vol.263 , pp. 802
    • Chalfie, M.1    Tu, Y.2    Euskirchen, G.3    Ward, W.W.4    Prasher, D.C.5
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    • note
    • In each of three replicates, about 40 eggs were hatched in 95 μl of M9 buffer plus 5 μl of pheromone extract (3).
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    • note
    • DR#271 and pRF4 plasmid (8) were microinjected into daf-7(n696).
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    • in preparation
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    • note
    • We thank C. Bargmann and J. White (neuron identification and discussion), M. Chalfie (gfp vector), J. Kramer (pRF4 plasmid), R. Barstead (cDNA library), H. R. Horvitz [daf-7(n896)], P. Hoppe and M. Estevez (help with microinjection), M. Estevez (preparing the Fig. 2 blot), M. Stanley and E. Norton (processing Fig. 3, A to D), V. Spate (γ source), D. Gems and C. Gunther (helpful discussions), and L. Hatley (manuscript processing). Supported by U.S. Department of Health and Human Services grant HD11239 to D.L.R.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.