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Volumn 289, Issue 5477, 2000, Pages 297-300

An inherited functional circadian clock in zebrafish embryos

Author keywords

[No Author keywords available]

Indexed keywords

MESSENGER RNA;

EID: 0034647717     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.289.5477.297     Document Type: Article
Times cited : (118)

References (33)
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    • note
    • A zebrafish Per3 polymerase chain reaction (PCR) probe was amplified with the primers 5′-CTGGG(C/ T)TA(C/T)CTICC(C/T)CAGGA-3′ and 5′-CAG(T/C)T(C/ G)ATCTGCTG(A/G)TAGGA-3′ and was used for screening a cDNA library of random-primed segmentation-stage zebrafish embryos and a cDNA library of oligo(dT)-primed maternal zebrafish embryos. A full-length and several overlapping independent cDNA clones were obtained and sequenced. The zebrafish Rev-erbα (NR1D1) probe was PCR amplified with the primers 5′-GT(T/G)GCITC(G/A)GG(C/A)TT(T/C)CA(C/ T)TA-3′ and 5′-GC(A/G)AA(C/T)TC(C/T)ACCACCTC(A/ C)CG-3′. The 17βHSD probe was used as a control (28).
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    • note
    • 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.
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    • note
    • The *AB zebrafish strain was kept at 28°C in a LD 14:10 cycle with lights on at 0900 hours (ZT 0). For most experiments, adults were crossed overnight, resulting in spawning and fertilization at about ZT 0, the next morning. Embryos were raised at 28°C in petri dishes containing 0.003% phenylthiourea to prevent pigmentation. Lighting conditions were either LD 14:10, LD 14:10 that was shifted B hours forward, LD 8:8, LL, or DD. Delayed development was obtained by keeping embryos at 23°C in LL. For desynchronizing development from the light/dark cycle, adults in LD 14:10 were crossed at ZT 3, and embryos fertilized at ZT 12 the same day were kept in LL. Embryos were fixed for 12 to 16 hours in 4% paraformaldehyde/phosphate-buffered saline at 4-hour intervals and stored in methanol. Unfertilized oocytes were obtained by squeezing mature females under anesthesia at 4-hour intervals.
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    • note
    • 32P-end-labeled oligonucleotide.
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    • note
    • We thank V. Heyer and G. Triqueneaux for technical assistance, O. Nkundwa and A. Karmim for maintenance of zebrafish, J.-M. Vanacker and M. Robinson-Rechavi for critical reading, and B. B. Rudkin for encouragement. This work was supported by funds from the INSERM, the CNRS, the Hôpital Universitaire de Strasbourg, the Association pour la Recherche sur le Cancer, and the Ligue Nationale de Lutte Contre le Cancer. F.D. is the recipient of a long-term postdoctoral fellowship from the Fondation pour la Recherche Médicale.


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.